galactomannan and Aspergillosis

Topics: Aspergillosis; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Intestinal Volvulus; Lymphoma, Large B-Cell, Diffuse; Male; Mannans; Nasopharyngeal Neoplasms; Neoplasm Recurrence, Local; Transplantation, Autologous; Young Adult

The spectrum of disease caused by Aspergillus spp. is dependent on the immune system of the host, and ranges from invasive aspergillosis (IA) to chronic pulmonary aspergillosis (CPA). Early and reliable diagnosis of Aspergillus disease is important to decrease associated morbidity and mortality. Areas covered: The following review will give an update on current diagnostic strategies for the diagnosis of IA and CPA. Expert commentary: Several new diagnostics for IA (including point-of-care tests) are now available to complement galactomannan testing. In particular, immunoPET/MRI imaging may be a promising approach for diagnosing IA in the near future. Notably, nearly all new biomarkers and tests for IA have been evaluated in the hematology setting only. Validation of biomarkers and tests is therefore needed for the increasing proportion of patients who develop IA outside the hematology setting. As an important first step, reliable definitions of IA are needed for non-hematology settings as clinical presentation and radiologic findings differ in these settings. CPA diagnosis is based on a combination of radiological findings in chest CT, mycological evidence (e.g. by the Aspergillus-specific IgG assay), exclusion of alternative diagnosis and chronicity. ([18F]FDG) PET/CT and immuno PET/MRI imaging are promising new imaging approaches.

Topics: Animals; Aspergillosis; Aspergillus; Biomarkers; Chronic Disease; Galactose; Humans; Magnetic Resonance Imaging; Mannans; Positron Emission Tomography Computed Tomography; Positron-Emission Tomography; Pulmonary Aspergillosis

Invasive aspergillosis (IA) is a life-threatening opportunistic mycosis that occurs in some people with a compromised immune system. The serum galactomannan enzyme-linked immunosorbent assay (ELISA) rapidly gained widespread acceptance as part of the diagnostic work-up of a patient suspected of IA. Due to its non-invasive nature, it can be used as a routine screening test. The ELISA can also be performed on bronchoalveolar lavage (BAL), allowing sampling of the immediate vicinity of the infection. The invasive nature of acquiring BAL, however, changes the role of the galactomannan test significantly, for example by precluding its use as a routine screening test.. To assess the diagnostic accuracy of galactomannan detection in BAL for the diagnosis of IA in people who are immunocompromised, at different cut-off values for test positivity, in accordance with the Cochrane Diagnostic Test Accuracy Handbook.. We searched three bibliographic databases including MEDLINE on 9 September 2016 for aspergillosis and galactomannan as text words and subject headings where appropriate. We checked reference lists of included studies for additional studies.. We included cohort studies that examined the accuracy of BAL galactomannan for the diagnosis of IA in immunocompromised patients if they used the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) classification as reference standard.. Two review authors assessed study quality and extracted data. Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) was used for quality assessment.. We included 17 studies in our review. All studies except one had a high risk of bias in two or more domains. The diagnostic performance of an optical density index (ODI) of 0.5 as cut-off value was reported in 12 studies (with 1123 patients). The estimated sensitivity was 0.88 (95% confidence interval (CI) 0.75 to 1.00) and specificity 0.81 (95% CI 0.71 to 0.91). The performance of an ODI of 1.0 as cut-off value could be determined in 11 studies (with 648 patients). The sensitivity was 0.78 (95% CI 0.61 to 0.95) and specificity 0.93 (95% CI 0.87 to 0.98). At a cut-off ODI of 1.5 or higher, the heterogeneity in specificity decreased significantly and was invariably >90%.. The optimal cut-off value depends on the local incidence and clinical pathway. At a prevalence of 12% a hypothetical population of 1000 patients will consist of 120 patients with IA. At a cut-off value of 0.5 14 patients with IA will be missed and there will be 167 patients incorrectly diagnosed with IA. If we use the test at a cut-off value of 1.0, we will miss 26 patients with IA. And there will be 62 patients incorrectly diagnosed with invasive aspergillosis. The populations and results were very heterogeneous. Therefore, interpretation and extrapolation of these results has to be performed with caution. A test result of 1.5 ODI or higher appears a strong indicator of IA.

Topics: Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunocompromised Host; Invasive Fungal Infections; Mannans; Randomized Controlled Trials as Topic; Sensitivity and Specificity

Topics: Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunocompromised Host; Invasive Fungal Infections; Mannans; Practice Guidelines as Topic; Sensitivity and Specificity

The serum galactomannan (GM) assay is used to diagnose invasive aspergillosis (IA). We conducted a systematic review and analysis to estimate the overall accuracy of the serum GM test for diagnosing pediatric IA.. A systematic literature review was conducted of all relevant studies published in PubMed and EMbase databases up to March 10, 2017. We selected and assessed articles that reported diagnostic data related to serum GM for diagnosis of pediatric IA. Pooled diagnostic odds ratios (DORs) and summary receiver operating characteristics (SROCs) were constructed with a cutoff value of 0.5. Additionally, pooled sensitivity (SEN), specificity (SPE), and positive and negative likelihood ratios (PLR and NLR, respectively) were estimated for summarizing overall test performance.. Seventeen studies were included in this systematic review. The total number of patients (age range 0-21 years old) was 1768, with 178 that had proven or probable IA. The pooled serum GM assay results, with a cutoff value of 0.5 for proven or probable IA, were DOR: 41.16 (95% confidence interval (CI) 21.48-78.86), SEN: 0.85 (95% CI 0.72-0.93), SPE: 0.88 (95% CI 0.80-0.93), PLR: 6.92 (95% CI 4.40-10.88), and NLR: 0.17 (95% CI 0.09-0.32). The SROC was 0.93.. Serum GM can be used to assist in diagnosis of proven or probable pediatric IA. However, serum GM test results should be interpreted in combination with clinical findings in pediatric IA cases, as the test results are not always sensitive or specific enough for pediatric IA.

Topics: Adolescent; Adult; Aspergillosis; Biomarkers; Child; Child, Preschool; Databases, Factual; Diagnostic Tests, Routine; Galactose; Humans; Infant; Infant, Newborn; Invasive Pulmonary Aspergillosis; Mannans; Meta-Analysis as Topic; Probability; ROC Curve; Sensitivity and Specificity; Young Adult

Clinical trials for invasive pulmonary aspergillosis, a potentially lethal mold infection, are complex investigations that require protracted time and extensive resources, delaying the development of new antifungal agents for this important disease. Areas covered: In this paper, the authors examine a novel approach to study invasive pulmonary aspergillosis in humans, with a focus on the potentials and pitfalls of surrogate end points such as galactomannan antigenemia to evaluate therapeutic response to novel compounds. Expert commentary: The authors believe the use of serum galactomannan as a primary end point in clinical trials may allow for development studies that could be accomplished with fewer resources. It is widely appreciated that serum galactomannan values strongly correlate with outcome of invasive aspergillosis in patients with hematologic cancer and may be applicable to other at-risk patient populations.

Topics: Antibodies, Fungal; Antifungal Agents; Aspergillosis; Biomarkers; Clinical Trials as Topic; Galactose; Humans; Mannans; Patient Care

Invasive mould infections (IMIs), such as invasive aspergillosis or mucormycosis, are a major cause of death in patients with haematological cancer and in patients receiving long-term immunosuppressive therapy. Early diagnosis and prompt initiation of antifungal therapy are crucial steps in the management of patients with IMI. The diagnosis of IMI remains a major challenge, with an increased spectrum of fungal pathogens and a diversity of clinical and radiological presentations within the expanding spectrum of immunocompromised hosts. Diagnosis is difficult to establish and is expressed on a scale of probability (proven, probable and possible). Imaging (CT scan), microbiological tools (direct examination, culture, PCR, fungal biomarkers) and histopathology are the pillars of the diagnostic work-up of IMI. None of the currently available diagnostic tests provides sufficient sensitivity and specificity alone, so the optimal approach relies on a combination of multiple diagnostic strategies, including imaging, fungal biomarkers (galactomannan and 1,3-β-d-glucan) and molecular tools. In recent years, the development of PCR for filamentous fungi (primarily Aspergillus or Mucorales) and the progress made in the standardization of fungal PCR technology, may lead to future advances in the field. The appropriate diagnostic approach for IMI should be individualized to each centre, taking into account the local epidemiology of IMI and the availability of diagnostic tests.

Topics: Aspergillosis; Aspergillus fumigatus; beta-Glucans; Early Diagnosis; Galactose; Hematologic Neoplasms; Humans; Immunocompromised Host; Immunosuppression Therapy; Invasive Fungal Infections; Magnetic Resonance Imaging; Mannans; Mucor; Mucormycosis; Organ Transplantation; Polymerase Chain Reaction; Positron Emission Tomography Computed Tomography; Tomography, X-Ray Computed

Infections caused by filamentous fungi represent a major burden in the ICU. Invasive aspergillosis is emerging in non-neutropenic individuals with predisposing conditions, e.g. corticosteroid treatment, chronic obstructive pulmonary disease, liver cirrhosis, solid organ cancer, HIV infection and transplantation. Diagnosis is challenging because the signs and symptoms are non-specific, and initiation of additional diagnostic examinations is often delayed because clinical suspicion is low. Isolation of an Aspergillus species from the respiratory tract in critically ill patients, and tests such as serum galactomannan, bronchoalveolar lavage 1-3-β-d-glucan and specific PCR should be interpreted with caution. ICU patients should start adequate antifungal therapy upon suspicion of invasive aspergillosis, without awaiting definitive proof. Voriconazole, and now isavuconazole, are the drugs of choice. Mucormycosis is a rare, but increasingly prevalent disease that occurs mainly in patients with uncontrolled diabetes mellitus, immunocompromised individuals or previously healthy patients with open wounds contaminated with Mucorales. A high proportion of cases are diagnosed in the ICU. Rapidly progressing necrotizing lesions in the rhino-sinusal area, the lungs or skin and soft tissues are the characteristic presentation. Confirmation of diagnosis is based on demonstration of tissue invasion by non-septate hyphae, and by new promising molecular techniques. Control of underlying predisposing conditions, rapid surgical resection and administration of liposomal amphotericin B are the main therapeutic actions, but new agents such as isavuconazole are a promising alternative. Patients with mucormycosis receive a substantial part of their care in ICUs and, despite advances in diagnosis and treatment, mortality remains very high.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; beta-Glucans; Critical Illness; Galactose; Humans; Immunocompromised Host; Intensive Care Units; Invasive Fungal Infections; Lung Diseases, Fungal; Mannans; Mucor; Mucormycosis; Nitriles; Opportunistic Infections; Pyridines; Respiratory System; Triazoles; Voriconazole

Although. Aspergillus spp infection is not the major cause of morbidity in Intensive Care Units (ICUs), mortality among patients treated for it is tremendous. Moreover, invasive aspergillosis (IA) is an independent risk factor of hospital costs and length of stay. The prevalence of this disease is inversely correlated with the immunocompetence of individuals; for instance, the incidence of IA among patients with leukemia is estimated as high as 12.7%. Although there is a significant improvement in the antifungal armamentarium, the appropriate treatment is still being given too late, mostly because of late diagnosis. As well as the diagnosis, the criteria for recognition of IA constitute a challenge.. The aim of this review, based on a case report, is to introduce the problem of poor diagnosis and treatment of IA, especially in the critical care settings. The presented scenario is an example which assists in showing the evidence-based medicine (EBM) approach to the treatment of fungal infections. Furthermore, to demonstrate the appropriate approach to diagnosis and treatment of invasive aspergillosis, the guidelines of The European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) are presented.. According to presented literature, Galactomannan assay enables early diagnosis and remains a specific and sensitive tool to diagnose Asppergillosis, both in serum and BAL fluid. The guidelines recommend voriconazole as a first line treatment in IA. Failure to detect and implement proper antifungal treatment may lead to fatal consequences, as in the presented case.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Critical Care; Fatal Outcome; Female; Galactose; Humans; Invasive Fungal Infections; Mannans; Middle Aged

Invasive fungal diseases are important clinical problems that are often complicated by severe illness and therefore the inability to use invasive measures to definitively diagnose the disease. Tests for a range of fungal biomarkers that do not require an invasive sample-collection procedure have been incorporated into adult clinical practice, but pediatric data and pediatric-specific recommendations for some of these diagnostic tools are lacking. In this review, we summarize the published literature and contemporary strategies for using the biomarkers galactomannan, (1→3)-β-d-glucan, Candida mannan antigen and anti-mannan antibody, and fungal polymerase chain reaction for diagnosing invasive fungal disease in children. Data on biomarker use in neonates and children with cancer, history of hematopoietic stem cell transplant, or primary immunodeficiency are included. Fungal biomarker tests performed on blood, other body fluids, or tissue specimens represent promising adjuncts to the diagnostic armamentarium in populations with a high prevalence of invasive fungal disease, but substantial gaps exist in the correct use and interpretation of these diagnostic tools in pediatric patients.

Topics: Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Biomarkers; Candida; Candidiasis; Child; DNA, Fungal; Galactose; Humans; Immunoassay; Invasive Fungal Infections; Mannans; Polymerase Chain Reaction

Fungi of the genus Aspergillus are ubiquitously present. Even though humans inhale Aspergillus spores daily under natural conditions, Aspergillus-associated pulmonary diseases only occur under special circumstances. Whether an Aspergillus-associated disease develops and which type of Aspergillus-associated disease develops depends on the constitution of the host. The spectrum of Aspergillus-associated pulmonary diseases ranges from allergic diseases, such as hypersensitivity pneumonitis to allergic infectious diseases, such as allergic bronchopulmonary aspergillosis (ABPA) and bronchocentric granulomatosis (BG) to infectious diseases, such as invasive (IA) or semi-invasive aspergillosis (SIA) and chronic pulmonary aspergillosis (CPA). Identification of Aspergillus spp. from sputum or bronchopulmonary secretions is not sufficient for a definitive diagnosis of Aspergillus-associated infections. The gold standard is the identification of Aspergillus spp. from lung tissue by culture or by histopathological methods; however, in clinical practice the decision to initiate antifungal therapy is more often based on immunological methods, such as the detection of Aspergillus-specific IgG antibodies from peripheral blood or galactomannan antigens from bronchoalveolar lavages. Acute IA or SIA infections have a high mortality and require immediate antifungal therapy. With rare exceptions CPA cannot be cured by medicinal therapy alone; however, active CPA can be brought into remission with antifungal therapy. Eradication of Aspergillus in CPA can as a rule only be successful using a combined antimycotic and surgical intervention.

Topics: Antibodies, Fungal; Antifungal Agents; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunoglobulin G; Invasive Pulmonary Aspergillosis; Lung; Lung Diseases, Fungal; Mannans; Respiratory System; Virulence

Galactomannan (GM) is a polysaccharide present in the cell wall of Aspergillus spp. that is released during growth of the organism. It has been successfully used to aide in the diagnosis of invasive aspergillosis allowing for earlier recognition of disease compared to conventional methods. Since its implementation in the clinic as a diagnostic tool, GM has been used in experimental models to measure therapeutic response. Several clinical studies describe the prognostic value of GM. Herein, we review the evidence supporting the utilization of GM antigen as a biomarker to measure response to systemic antifungal therapy.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Biomarkers; Galactose; Humans; Mannans; Prognosis

The clinical utility of bronchoalveolar lavage (BAL) fluid galactomannan (GM) for the early diagnosis of invasive aspergillosis (IA) varies widely across studies mainly due to heterogeneity of the studied populations.. We conducted a systematic review and meta-analysis of 16 studies involving 783 adults with hematological malignancies to derive summary estimates of the overall accuracy of BAL-GM for diagnosing IA.. Summary estimates of BAL-GM using an optical density (OD) index cutoff value of 1.5 for proven and probable IA were: sensitivity 0.92 (95% CI = 0.48-0.99), specificity 0.98 (95% CI = 0.78-1.00), positive likelihood ratio 53.7 (95% CI = 3.7-771.8), and negative likelihood ratio 0.08 (95% CI = 0.01-0.83). Comparing serum GM and Aspergillus PCR testing on BAL fluid, BAL-GM conferred greater sensitivity, but lower specificity than the serum GM test, and similar specificity as the PCR assay. The use of BAL-GM with serum GM or BAL-PCR tests increased the sensitivity moderately when a positive result was defined by either assay.. GM quantification in BAL fluid at an OD index cutoff value of 1.5 has excellent sensitivity and specificity to assist clinical decision-making in confirming or excluding a diagnosis of IA when results are interpreted with clinical findings. Additional research investigating the effects of antifungal agents, optimal timing and processing of BAL sampling are needed to improve the diagnostic accuracy of BAL-GM testing.

Topics: Adult; Aspergillosis; Bronchoalveolar Lavage Fluid; Galactose; Hematologic Neoplasms; Humans; Mannans; Polymerase Chain Reaction; Sensitivity and Specificity

Invasive aspergillosis is an infection with high morbidity and mortality that affects mostly immunocompromised individuals. Early identification and targeted treatment of the infection is essential to improve survival of affected patients. The purpose of our review is to highlight the most recent developments in diagnosis and screening for invasive aspergillosis (IA) along with the challenges associated with the development and validation of novel diagnostic approaches.. Ovid MEDLINE and The Cochrane library were searched for studies that evaluated serologic, molecular and novel methodologies for the diagnosis of IA.. Traditional diagnostic approaches, such as histopathology and culture, are still considered the gold standard but lack sufficient sensitivity. Newer serologic techniques, such as galactomannan (GM) and beta-glucan, have already been incorporated into clinical guidelines, but recent evidence suggests that their performance might be limited in certain clinical settings. Molecular methods, such as the Aspergillus spp. polymerase chain reaction (PCR), have not yet found their place in clinical practice mainly due to lack of standardization. Novel methodologies, such as volatile organic compound detection and lateral flow devices, have recently been developed and promise noninvasive and rapid diagnosis of aspergillosis, while diagnostic algorithms that incorporate both GM and PCR have proven to be effective in early randomized trials as screening methods and can reduce the use of antifungal agents.. Diagnosis of IA remains challenging. Novel methodologies and the standardization of GM and PCR might provide more reliable diagnostic tools in the future.

Topics: Aspergillosis; beta-Glucans; Early Diagnosis; Galactose; Humans; Mannans; Real-Time Polymerase Chain Reaction; Volatile Organic Compounds

Aspergillus polymerase chain reaction (PCR) was excluded from the European Organisation for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) definitions of invasive fungal disease because of limited standardization and validation. The definitions are being revised.. A systematic literature review was performed to identify analytical and clinical information available on inclusion of galactomannan enzyme immunoassay (GM-EIA) (2002) and β-d-glucan (2008), providing a minimal threshold when considering PCR. Categorical parameters and statistical performance were compared.. When incorporated, GM-EIA and β-d-glucan sensitivities and specificities for diagnosing invasive aspergillosis were 81.6% and 91.6%, and 76.9% and 89.4%, respectively. Aspergillus PCR has similar sensitivity and specificity (76.8%-88.0% and 75.0%-94.5%, respectively) and comparable utility. Methodological recommendations and commercial PCR assays assist standardization. Although all tests have limitations, currently, PCR is the only test with independent quality control.. We propose that there is sufficient evidence that is at least equivalent to that used to include GM-EIA and β-d-glucan testing, and that PCR is now mature enough for inclusion in the EORTC/MSG definitions.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Galactose; Humans; Immunoenzyme Techniques; Mannans; Polymerase Chain Reaction; Quality Control; Sensitivity and Specificity

Screening of high-risk patients for invasive aspergillosis (IA) has the potential to decrease the use of empiric antifungal agents. However, the performance of different screening methods has not been studied.. We performed a meta-analysis of published studies to assess the diagnostic performance of galactomannan (GM) and polymerase chain reaction (PCR) as weekly screening tests in high-risk populations. The sensitivity and specificity of 6 approaches combining GM and PCR were estimated using the bivariate model.. Thirteen studies with 1670 patients met our inclusion criteria. Single positive test results had modest sensitivity and specificity for screening (respectively, 92% and 90% for GM; 84% and 76% for PCR). The screening approach with the highest sensitivity was the one that used at least 1 GM- or PCR-positive result to define a positive episode, achieving a sensitivity of 99%, significantly higher than any single test (P = .0018 compared with GM and P < .0001 compared with PCR). Meanwhile, when both GM and PCR were positive for the same patient, the specificity increased to 98%, which was not significantly different compared to the specificity of at least 2 positive GM (95%, P = .56 for the comparison) or PCR results (93%, P = .07 for the comparison).. When screening high-risk patients for IA with GM and PCR tests, the absence of any positive test can obviate the need for antifungal agents with a negative predictive value of 100%, whereas the presence of at least 2 positive results is highly suggestive of an active infection with a positive predictive value of 88%.

Topics: Aspergillosis; Aspergillus; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Mass Screening; Polymerase Chain Reaction; Risk Factors; Sensitivity and Specificity

An established diagnosis of invasive aspergillus is seldom achieved premortem. Conventional laboratory diagnostic methods such as culture and microscopy, although very useful when positive, are insensitive and time-consuming, resulting in late diagnosis and treatment and contributing to high mortality rates. As a result, routine antifungal prophylaxis and early empirical treatment have been recommended. The use of sensitive and rapid non-culture-based diagnostic assays for the detection of Aspergillus antigens (using commercially available tests to detect galactomannan and 1, 3 β-D-glucan) or detection of genomic DNA sequences may allow a shift in emphasis from empirical to preemptive therapy, especially when substantiated by suggestive radiological findings. These new tools may be used to confirm a presumed diagnosis of invasive aspergillosis, or, when used to screen high-risk patients, may identify an infection at an early stage of disease. Their excellent negative predictive value should convince clinicians to withhold antifungal therapy in patients with no other signs of fungal disease. On the other hand, consecutive positive results should at least trigger a complete diagnostic workup. This article will review the diagnostic utility as well as the pitfalls of using these non-culture-based tools for diagnosing invasive aspergillosis.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; beta-Glucans; Bronchoalveolar Lavage Fluid; Galactose; Humans; Mannans; Proteoglycans; Triazoles; Voriconazole

Invasive aspergillosis is the most common life-threatening opportunistic invasive mycosis in immunocompromised patients. A test for invasive aspergillosis should neither be too invasive nor too great a burden for the already weakened patient. The serum galactomannan enzyme-linked immunosorbent assay (ELISA) seems to have the potential to meet both requirements.. To obtain summary estimates of the diagnostic accuracy of galactomannan detection in serum for the diagnosis of invasive aspergillosis.. We searched MEDLINE, EMBASE and Web of Science with both MeSH terms and text words for both aspergillosis and the sandwich ELISA. We checked the reference lists of included studies and review articles for additional studies. We conducted the searches in February 2014.. We included cross-sectional studies, case-control designs and consecutive series of patients assessing the diagnostic accuracy of galactomannan detection for the diagnosis of invasive aspergillosis in patients with neutropenia or patients whose neutrophils are functionally compromised. The reference standard was composed of the criteria given by the European Organization for Research and Treatment of Cancer (EORTC) and the Mycoses Study Group (MSG).. Two review authors independently assessed quality and extracted data. We carried out meta-analysis using the bivariate method. We investigated sources of heterogeneity by adding potential sources of heterogeneity to the model as covariates.. We included 54 studies in the review (50 in the meta-analyses), containing 5660 patients, of whom 586 had proven or probable invasive aspergillosis. When using an optical density index (ODI) of 0.5 as a cut-off value, the sensitivity of the test was 82% (73% to 90%) and the specificity was 81% (72% to 90%). At a cut-off value of 1.0 ODI, the sensitivity was 72% (65% to 80%) and the specificity was 88% (84% to 92%). At a cut-off value of 1.5 ODI, the sensitivity was 61% (47% to 75%) and the specificity was 93% (89% to 97%). None of the potential sources of heterogeneity had a statistically significant effect on either sensitivity or specificity.. If we used the test at a cut-off value of 0.5 ODI in a population of 100 patients with a disease prevalence of 9% (overall median prevalence), two patients who have invasive aspergillosis would be missed (sensitivity 82%, 18% false negatives), and 17 patients would be treated unnecessarily or referred unnecessarily for further testing (specificity 81%, 19% false negatives). If we used the test at a cut-off value of 1.5 in the same population, that would mean that four invasive aspergillosis patients would be missed (sensitivity 61%, 39% false negatives), and six patients would be treated or referred for further testing unnecessarily (specificity 93%, 7% false negatives). These numbers should, however, be interpreted with caution because the results were very heterogeneous.

Topics: Aspergillosis; Biomarkers; Galactose; Humans; Immunocompromised Host; Mannans; Opportunistic Infections; Sensitivity and Specificity

The biomarkers galactomannan and 1,3-β-d-glucan have been well studied over the past years and are gaining a role in the diagnosis of invasive fungal infections. Although not as well studied until recently, molecular methods for the diagnosis of invasive fungal infection are also being evaluated. Outcomes data for molecular testing are expanding, but have not yet provided enough evidence for inclusion of molecular diagnostics in formal clinical guidelines. Lack of standardization and validation of the various molecular assays and platforms has hindered their widespread acceptance in the evaluation of invasive fungal infections, although the future is promising.

Topics: Aspergillosis; beta-Glucans; Biomarkers; Bronchoalveolar Lavage Fluid; Candidiasis; False Positive Reactions; Galactose; Humans; Mannans; Pathology, Molecular; Polymerase Chain Reaction; Predictive Value of Tests; Sugar Alcohols

Topics: Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Biomarkers; Diagnosis, Differential; Early Diagnosis; Galactose; Humans; Immunocompromised Host; Mannans; Molecular Diagnostic Techniques; Proteoglycans; Reagent Kits, Diagnostic; Tomography, X-Ray Computed

Invasive aspergillosis, chronic pulmonary aspergillosis and allergic bronchopulmonary aspergillosis are the clinical forms of aspergillosis. Although there is a great number of Aspergillus species, Aspergillus fumigatus-complex is the more frequent aetiological agent, regardless of clinical form or baseline condition. The increase in immunosuppressive agents and the higher use of corticosteroids in chronic obstructive pulmonary disease have led to aspergillosis becoming more prominent in recent years. Galactomannan detection and radiological diagnostic images complement the limitations of microbiology cultures in these patients. Voriconazole and liposomal amphotericin B are the gold standard in patients requiring therapy, and posaconazole, itraconazole, caspofungin and other echinocandins are effective alternatives. The prognosis depends of clinical forms and characteristics of the host, but it is particularly poor in the disseminated invasive forms.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Cross Infection; Drug Resistance, Multiple, Fungal; Endocarditis; Endophthalmitis; Fungemia; Galactose; Humans; Immunocompromised Host; Mannans; Neuroaspergillosis; Postoperative Complications; Pulmonary Aspergillosis; Radiography; Risk Factors; Salvage Therapy; Species Specificity; Vulnerable Populations

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Breath Tests; DNA, Fungal; Early Diagnosis; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Leukemia; Lung; Mannans; Neutropenia; Organ Transplantation; Transplantation Immunology

Invasive fungal infections (IFIs) are difficult to diagnose, especially early in the course of infection when antifungal therapy is most effective. There are two commercially available biomarker assays useful for detection of the IFIs most commonly seen in patients with hematologic malignancies, the galactomannan and beta glucan assays. The former is specific for aspergillosis, the latter positive for not only Aspergillus and Candida species, but several other clinically relevant fungal pathogens as well. Both have good assay performance characteristics, provide rapid test results, are widely available, can be assayed non-invasively, and are positive early in the course of infection, often before onset of signs and symptoms of infection. Adoption of these assays into clinical practice has led to reduced need to perform invasive procedures to obtain deep tissue to establish the diagnosis of invasive fungal infections. Improved survival rates from aspergillosis are, in part, due to earlier detection of infection and earlier therapy.

Topics: Acute Disease; Aspergillosis; Aspergillus; beta-Glucans; Biomarkers; Candida; Candidiasis; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Leukemia; Mannans; Transplantation, Homologous

Early mycological detection of Aspergillus species is the cornerstone for a prompt diagnosis, appropriate treatment strategies, and improved survival of patients with invasive aspergillosis (IA), irrespective of age. However, the currently available laboratory tests for the diagnosis of IA include culture with direct microscopy, histology, and antigenic markers, such as galactomannan and β-1, 3-d-glucan, Aspergillus spp. DNA detection by PCR, and imaging studies, such as high-resolution CT scan. However, all need further validation, especially in children. In this review we focus on the diagnosis of IA emphasizing the current perspectives, difficulties in interpretation, and the need of further evaluation from a pediatric point of view.

Topics: Aspergillosis; Aspergillus; Biomarkers; Child; DNA, Fungal; Galactose; Humans; Mannans; Polymerase Chain Reaction

Invasive aspergillosis (IA) is a major cause of mortality in patients with hematological malignancies, due largely to the inability of traditional culture and biopsy methods to make an early or accurate diagnosis. Diagnostic accuracy studies suggest that Aspergillus galactomannan (GM) enzyme immunoassay (ELISA) and Aspergillus PCR-based methods may overcome these limitations, but their impact on patient outcomes should be evaluated in a diagnostic randomized controlled trial (D-RCT). This article describes the methodology of a D-RCT which compares a new pre-emptive strategy (GM-ELISA- and Aspergillus PCR-driven antifungal therapy) with the standard fever-driven empiric antifungal treatment strategy. Issues including primary end-point and patient selection, duration of screening, choice of tests for the pre-emptive strategy, antifungal prophylaxis and bias control, which were considered in the design of the trial, are discussed. We suggest that the template presented herein is considered by researchers when evaluating the utility of new diagnostic tests (ClinicalTrials.gov number, NCT00163722).

Topics: Antifungal Agents; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Hematologic Neoplasms; Humans; Mannans; Polymerase Chain Reaction; Randomized Controlled Trials as Topic; Research Design; Risk Factors

Although invasive fungal infections (IFIs) are relatively rare, they are important causes of morbidity and mortality in immunocompromised pediatric patients. Early and precise diagnosis of IFI is important to allow antifungal treatment to be started in time and to reduce the unnecessary use of toxic antifungal agents. Although traditional approaches such as direct microscopic examination, histopathological evaluation and cultivation are still gold standard, the diagnosis of IFI is generally difficult because of inadequate sensitivity and specificity with these tests. Commercial systems detecting the Aspergillus cell wall antigen galactomannan and 1,3-β-D-glucan are seen as the most convenient nonculture methods for the diagnosis of the IFI and monitoring of antifungal treatment. Several molecular methods have been described for the diagnosis of opportunistic mycoses. However, they have not been standardized and have only been used in experimental studies.

Topics: Aspergillosis; Aspergillus; beta-Glucans; Candida; Candidiasis, Invasive; Child; Culture Techniques; Diagnostic Techniques and Procedures; Early Diagnosis; Galactose; Humans; Mannans; Sensitivity and Specificity

The (1→3)-β-D-Glucan (BG) assay has been approved for diagnosing invasive fungal disease (IFD). However, the test performance has been variable. We conducted a meta-analysis to determine the overall accuracy of BG assay for diagnosing IFD.. The sensitivity, specificity, and positive and negative likelihood ratios (PLR and NLR, respectively) of BG for diagnosing IFD were pooled using a bivariate meta-analysis. We also performed subgroup analyses.. Twelve reports, including 15 studies, were included for the analysis (proven and probable IFD vs possible or no IFD). The sensitivity, specificity, PLR and NLR were 0.76 (95% CI, 0.67-0.83), 0.85 (95% CI, 0.73-0.92), 5.05 (95% CI, 2.71-9.43), and 0.28 (95% CI, 0.20-0.39), respectively. Subgroup analyses showed that the BG assay had higher specificities for patients with hematological disorders and a positive BG result with two consecutive samples. The combination of galactomannan and BG increased the specificity value to 0.98 (95% CI, 0.95-0.99) for diagnosing invasive aspergillosis.. Serum BG determination is clinically useful for diagnosing IFD in at-risk patients, especially for hematology patients. The combination of galactomannan and BG was sufficient for diagnosing invasive aspergillosis. Since the BG assay is not absolutely sensitive and specific for IFD, the BG results should be interpreted in parallel with clinical findings.

Topics: Aspergillosis; beta-Glucans; Biomarkers; Candidiasis, Invasive; Galactose; Humans; Likelihood Functions; Linear Models; Mannans; Mycoses; Proteoglycans; ROC Curve; Sensitivity and Specificity

Diseases caused by Aspergillus spp. are difficult to diagnose and thus require supplementary serological assays. This is the result of a selective review of the relevant literature with special regard to recent guidelines. In addition to conventional diagnostic tools (radiology, microscopy, culture) the measurement of the following serological markers is recommended, depending on the clinical type of aspergillosis: Invasive and chronic necrotising aspergillosis: Aspergillus-galactomannan antigen. Test format: EIA using the rat MAb EB-A2. Cut-off 0.5 (index). Monitoring of high risk patients: Twice weekly. Aspergillus-IgG (test format EIA) as confirmatory assay after recovery of the leukocyte function under therapy. Aspergilloma: Aspergillus IgG. Test format: EIA. Allergical aspergillosis: Aspergillus IgE. Test format: RAST. Galactomannan antigen detection rates high in the diagnosis of invasive aspergillosis. The evaluation of Aspergillus nucleic acid amplification assays is pending.

Topics: Antibodies, Fungal; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Humans; Immunoenzyme Techniques; Immunoglobulin E; Immunoglobulin G; Mannans; Practice Guidelines as Topic; Serologic Tests

Invasive aspergillosis is a major cause of mortality in allogeneic bone marrow transplant recipients and patients treated for blood malignancies. The diagnostic tools, treatments and preventive strategies, essentially developed for neutropaenic patients, have not been assessed in populations whose immune systems are considered to be competent.. Beside the standard picture of chronic Aspergillus infection, the incidence of invasive aspergillosis is increasing in non neutropaenic patients, such as those with chronic lung diseases or systemic disease treated with long-term immunosuppressive drugs and solid organ transplant recipients. This study reviews the specific features of invasive aspergillosis in non neutropaenic subjects (NNS) and discusses the value of the diagnostic tools and treatment in this population.. A better understanding of the pathophysiology and the epidemiological characteristics of invasive aspergillosis would provide a means of adapting the staging and classification of the disease for NNS.. Invasive aspergillosis is under diagnosed in NNS who may already be colonised when they receive immunosuppressive treatment; this can lead to an adverse outcome in patients who are considered to be a moderate risk population.

Topics: AIDS-Related Opportunistic Infections; Antibodies, Fungal; Antifungal Agents; Aspergillosis; Aspergillus; Chronic Disease; Fungemia; Galactose; Humans; Immunocompromised Host; Immunosuppressive Agents; Lung Diseases; Mannans; Neutropenia; Organ Transplantation; Postoperative Complications; Pulmonary Aspergillosis; Radiography; Respiratory System; Risk Factors; Wound Infection

To assess the value of galactomannan (GM) double-direct sandwich enzyme-linked immunosorbent assay (ELISA) in the diagnosis of invasive aspergillosis (IA).. A search in MEDLINE, EMbase, OVID, CBMdisc and CHKD from Jan. 1991 to Dec. 2008 was conducted to collect all articles about diagnostic tests of serum GM detection. Then the methodological quality was assessed by QUADAS-items, sources of heterogeneity investigated, pooled effect quantities evaluated, and meta-analysis studies, SROC curves, and subgroup analysis performed.. Thirty-six articles with a population of 4959 patients were included. The average prevalence of IA was 10%(532/4959). Our meta-analysis reported a median heterogeneity (I(2) = 48.6%, P < 0.05), with a pooled DOR value of 19.10 (95%CI 12.67 - 28.79), a pooled sensitivity of 0.66 (95%CI 0.61 - 0.70), a pooled specificity of 0.90 (95%CI 0.89 - 0.90), a pooled positive likelihood ratio of 5.48 (95%CI 4.27 - 7.02), a pooled negative likelihood ratio of 0.38 (95%CI 0.29 - 0.50), and an area under curve of SROC 0.88. The rate of underdiagnosis of serum GM detection was 34% (168/490) and the rate of misdiagnosis was 10% (466/4469). With a rise in the cut-off value the sensitivity of GM test decreased and specificity increased. Two consecutive positive tests decreased the sensitivity but increased the specificity. Age had no significant effect on the diagnosis by GM test. Both antifungal prophylaxis and antifungal therapy had no significant effect on sensitivity and specificity of GM test for IA diagnosis.. Serum GM detection is an effective diagnostic tool for invasive aspergillosis in high-risk populations.

Topics: Antigens, Fungal; Aspergillosis; Galactose; Humans; Mannans; Sensitivity and Specificity

Topics: Antifungal Agents; Aspergillosis; beta-Glucans; Biomarkers; Candidiasis; Galactose; Hematologic Diseases; Humans; Immunocompromised Host; Lung Diseases, Fungal; Mannans; Molecular Diagnostic Techniques; Mycoses

Invasive aspergillosis is a common cause of morbidity and mortality in hematopoietic stem cells transplant recipients. Owing to its intrinsic high mortality rate, early diagnosis and treatment are critical. This review will therefore address the most important recent advances in diagnosing, preventing and treating invasive aspergillosis in hematopoietic stem cells transplant.. The present review will focus on therapeutic and prophylactic aspects, with particular regard to clinical use of drugs other than voriconazole (which has a well known and consolidated role for first-line therapy), combination therapy and prophylactic regimens, particularly with posaconazole. This review will also briefly deal with the clinical role of diagnostic tests such as the detection of galactomannan in body fluids other than blood, beta-D-glucan in serum and fungal DNA by PCR in body fluids.. Galactomannan antigen detection is a rather reliable diagnostic test for invasive aspergillosis, particularly when a lower threshold of sensitivity is used. PCR is still to be validated. Liposomal amphotericin B at 3 mg/kg per day showed a similar efficacy in invasive aspergillosis as reported for voriconazole. Therapeutic drug monitoring of Aspergillus-active azoles should be implemented whenever possible in order to maximize the antifungal effect and minimize toxicity. Posaconazole showed to be active in prophylaxis, though its effectiveness in the global patient population is still controversial.

Topics: Antifungal Agents; Aspergillosis; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans

Invasive aspergillosis (IA) is the most common life-threatening opportunistic invasive mycosis in immunocompromized patients. A test for IA needs to be not too invasive and not too big a burden for the already weakened patient. The serum galactomannan ELISA seems to have potential for both requirements.. To obtain summary estimates of the diagnostic accuracy of galactomannan detection in serum for the diagnosis of IA.. We searched MEDLINE, EMBASE and Web of Science with both Medical Headings and text words for both aspergillosis and the sandwich ELISA. We checked reference lists of included studies and review articles for additional studies.. Cross-sectional studies, case-control designs and consecutive series of patients assessing the diagnostic accuracy of galactomannan detection for the diagnosis of IA in patients with neutropenia or patients whose neutrophils are functionally compromised were included. The reference standard was composed of the criteria given by the European Organization for Research and Treatment of Cancer (EORTC) and the Mycoses Study Group (MSG).. Two review authors independently assessed quality and extracted data. Thirty studies were included in the meta-analyses, with a median prevalence of IA (proven or probable) of 7.7%. Seven of these (901 patients) reported results for an Optical Density Index (ODI) of 0.5 as cut-off value. The overall sensitivity in these studies was 78% (61% to 89%) and overall specificity was 81% (72% to 88%). Twelve studies (1744 patients) reported the results for cut-off value of 1.0 ODI, overall sensitivity was 75% (59% to 86%) and mean specificity 91% (84% to 95%). Seventeen studies (2600 patients) reported the results for cut-off value 1.5 ODI, sensitivity was 64% (50% to 77%) and mean specificity 95% (91% to 97%).. At a cut-off value 0.5 ODI in a population of 100 patients with a disease prevalence of 8% (overall median prevalence), 2 patients who have IA, will be missed (sensitivity 78%, 22% false negatives), and 17 patients will be treated or further referred unnecessarily (specificity of 81%, 19% false negatives). If we use the test at cut-off value 1.5 in the same population, that will mean that 3 IA patients will be missed (sensitivity 64%, 36% false negatives) and 5 patients will be treated or referred unnecessarily (specificity of 95%, 5% false negatives). These numbers should however be interpreted with caution, because the results were very heterogeneous.

Topics: Aspergillosis; Biomarkers; Galactose; Humans; Immunocompromised Host; Mannans; Opportunistic Infections; Randomized Controlled Trials as Topic; Sensitivity and Specificity

In the last few years, major advances in the treatment of transplant recipients, with hemato-oncological diseases or admitted to the intensive care unit, has been accompanied by an increase in classical fungal infections and by the emergence of uncommon fungal infections. Despite the development of new diagnostic techniques such as galactomannan detection and the availability of new antifungal agents, these opportunistic infections continue to pose a diagnostic challenge, prolong length of hospital stay, and increase costs. In addition, mortality from these infections is high. The present chapter provides a brief review of the epidemiology of these infections, diagnostic advances, and the new antifungal agents that have been developed in the last few years.

Topics: Anidulafungin; Antifungal Agents; Aspergillosis; beta-Glucans; Candidiasis; Clinical Trials as Topic; Critical Care; Diabetes Complications; Disease Susceptibility; Echinocandins; Fungemia; Galactose; Hematologic Diseases; Humans; Immunocompromised Host; Mannans; Meta-Analysis as Topic; Mycoses; Neoplasms; Opportunistic Infections

Invasive aspergillosis is a serious and often fatal infection in patients who are neutropenic or have undergone solid organ or stem cell transplantation. Delayed diagnosis and therapy may lead to poor outcomes. Diagnosis may be facilitated by a test for galactomannan antigen detection using an enzyme immunoassay. Other rapid methods for diagnosis include (1-->3)-beta-D: -glucan determination and polymerase chain reaction. The sensitivity and specificity of galactomannan antigenemia testing in serum and bronchoalveolar lavage specimens are high in patients with hematological malignancy, neutropenia, and receipt of stem-cell transplants. False positivity can be seen with concomitant administration of some antibiotics and infection by fungi other than Aspergillus.

Topics: Antigens, Fungal; Aspergillosis; Galactose; Humans; Immunoenzyme Techniques; Mannans

Invasive aspergillosis (IA) is an important infectious complication in allogeneic stem cell transplant (SCT) recipients. Diagnosis of IA has been difficult and often delayed and treatment outcome has been poor, with mortality rates up to 80%. This review summarizes recent developments in this field. There are indications that the incidence of IA may be decreasing due to multiple factors including better understanding of pathogenesis of IA, earlier diagnosis, and various prophylactic and preventive strategies. Recently posaconazole has shown to be effective in reducing the risk of IA in patients treated for graft-versus-host disease (GVHD). Early use of high-resolution thoracic computed tomography assisted with complimentary methods including bronchoalveolar lavage and serum galactomannan determinations are useful in early diagnosis. Our treatment armamentarium against IA has broadened significantly during the last years and there are some indications of improved outcome more recently. On the other hand, increasing use of blood progenitor grafts instead of marrow with higher risk of chronic GVHD, increasing age of SCT recipients, and wide use of donor lymphocyte infusions for treatment of minimal residual disease or relapse may affect to the opposite direction. Despite some promises and improvements, IA will continue to remain a challenge in the upcoming years.

Topics: Antifungal Agents; Aspergillosis; Bronchoalveolar Lavage Fluid; Galactose; Graft vs Host Disease; Hematopoietic Stem Cell Transplantation; Humans; Immunosuppression Therapy; Leukemia, Myeloid, Acute; Mannans; Polymerase Chain Reaction; Risk Factors; Transplantation, Homologous; Treatment Outcome

In high-risk patient cohorts, such as patients after solid-organ or allogeneic stem-cell transplantation, or patients with acute leukaemia, early diagnosis of invasive fungal infections (IFIs) is essential, as delayed or missing diagnosis of IFI results in increasing rates of mortality. However, diagnosis of most IFIs, especially of invasive aspergillosis, is difficult because classic tests have low sensitivity and specificity, and radiology often provides non-specific and transient results. The limited sensitivity and specificity of conventional assays for the detection of IFI and the growing number of immunocompromised patients who are at risk for opportunistic fungal infections have led to the development of new assays. These methods include antigen detection systems, such as ELISAs, and different molecular methods (PCR assays). Serological tests, such as the detection of the carbohydrate galactomannan, are standardised and commercially available. However, they still need to be evaluated in large patient cohorts, especially children. The benefit of antibody detection remains unclear if patients are under immune suppression or are heavily colonised but not infected. A range of different PCR assays (conventional, nested, real-time) have been developed, targeting different gene regions (cytochrome P450, heat-shock proteins, 18S, 5.8S, 28S, internal transcribed spacer), including a variety of amplicon detection methods, such as gel electrophoresis, hybridisation with specific probes, ELISA and restriction fragment length polymorphism. These molecular assays provide high potential in terms of sensitivity and specificity, but vary widely in their feasibility and up to now have not been standardised. Taken together, new non-culture-based diagnostic assays are appropriate as simple and rapid screening tests with high sensitivities and quick turnaround times. Thus, they might help to reduce empirical antifungal therapy and might be valuable tools to allow early initiation and monitoring of pre-emptive antifungal therapy. In this review, we assess the performance of a variety of non-culture-based tests for the detection of IFI in high-risk haematological patients, with emphasis on the impact of the assays on different management strategies.

Topics: Aspergillosis; Aspergillus; beta-Glucans; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Immunocompromised Host; Mannans; Polymerase Chain Reaction; Sensitivity and Specificity; Tomography, X-Ray Computed

Invasive aspergillosis is a serious and lethal infection among immunocompromised patients, with reported mortality rates as high as 74-92%. The high mortality is related to the severe immunosuppression experienced by these patients as well as the difficulties for physicians in arriving at a timely diagnosis. Definitive diagnostic procedures (tissue biopsy for histopathology and culture) are often precluded by severe cytopenias and coagulation abnormalities. The development of minimally invasive, nonculture diagnostic methods is a major advance in the early diagnosis of invasive aspergillosis. Galactomannan is a heteropolysaccharide (mannan core and side residues of galactofuranosyl units) present in the cell wall of Aspergillus spp. The double sandwich enzyme immunoassay, which detects galactomannan in serum samples, has been available in Europe for almost a decade and in the USA since May 2003, for the diagnosis of invasive aspergillosis. However, availability of the double galactomannan enzyme immunoassay is center variable in the USA and, although its analytical performance in the diagnosis of invasive aspergillosis is well documented, its routine use in clinical practice is limited. As an adjunct in the diagnosis and management of invasive aspergillosis, incorporation of the galactomannan enzyme immunoassay into clinical trials will help to further define its role.

Topics: Animals; Antigens, Fungal; Aspergillosis; Disease Models, Animal; Galactose; Humans; Immunohistochemistry; Mannans; Molecular Weight

Recognition and treatment of invasive fungal infections in acute leukemia and hematopoietic stem cell transplant patients are important clinical challenges. New diagnostic tools, such as fungal serologic assays and high-resolution CT scans, offer the hope for earlier initiation of antifungal therapy and improved treatment results. New antifungal agents offer choices that in some cases are less toxic than older drugs and in other cases are more efficacious. Combining the new diagnostic tools with new drugs, novel strategies are being evaluated to change our approaches to these deadly infections.

Topics: Antibiotic Prophylaxis; Antifungal Agents; Aspergillosis; Candidiasis; Clinical Trials as Topic; Galactose; Glucans; Hematopoietic Stem Cell Transplantation; Humans; Leukemia; Mannans

A reliable diagnosis of invasive aspergillosis in patients with haematological malignancies is seldom achieved antemortem. Conventional laboratory diagnostic methods are insensitive and time-consuming, resulting in late diagnosis and treatment and contributing to unacceptably high mortality. As a result, routine antifungal prophylaxis and early empirical treatment have been recommended. However, overtreatment associated with these strategies results in increased toxicity and cost. The use of sensitive and rapid non-culture-based diagnostic assays, such as detection of Aspergillus antigens (galactomannan, beta-D-glucan) or detection of genomic DNA sequences may allow a shift in emphasis from empirical to pre-emptive therapy, especially when substantiated by suggestive radiological findings. These new tools may be used to confirm a presumed diagnosis of invasive aspergillosis, or, when used to screen high-risk patients, may identify an infection at the early stage of disease. The excellent negative predictive value of these assays should convince clinicians to withhold antifungal therapy in persistently febrile neutropenic patients with no other signs of fungal infection. On the other hand, consecutive positive results in a high-risk population should at least trigger a complete diagnostic work-up. This review will focus on the diagnostic utility as well as on the pitfalls of serial screening for the presence of circulating fungal antigens in haematology patients.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Galactose; Hematologic Neoplasms; Humans; Immunoassay; Mannans; Proteoglycans; Serologic Tests

The diagnosis of invasive fungal infections (IFI) remains a challenge, particularly for diseases caused by filamentous fungi such as Aspergillus species. Unfortunately, many patients affected by these conditions are not identified before autopsy. Therefore, there is a need for new diagnostic methods for IFI. Galactomannan is a soluble antigen released during hyphal growth in tissues. A commercially available sandwich ELISA assay that detects galactomannan has been used in Europe for many years and is now approved for use in the USA. The test has an excellent negative predictive value in the detection of invasive aspergillosis (IA) in high-risk patients. In addition, it is more sensitive than culture and allows IA to be diagnosed before clinical manifestations occur. However, false-negative and false-positive results in certain populations are the main limitations to its use. The purpose of this review is to summarize the current knowledge about galactomannan testing in patients at risk for IA.

Topics: Animals; Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; False Negative Reactions; False Positive Reactions; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Mice; Organ Transplantation; Rabbits; Sensitivity and Specificity

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Humans; Mannans

Native valve endocarditis caused by Aspergillus spp. is an uncommon disease with a high mortality rate. Generally, Aspergillus is isolated from affected valve in post-mortem or biopsy specimens. However, its isolation from blood cultures is exceedingly rare. We report a case of fungal endocarditis in a native mitral valve with the isolation of Aspergillus fumigatus both in valve vegetation and in blood culture bottles. The patient underwent valve replacement and antifungal treatment with voriconazole and caspofungin, but he died on post-operative day 45 with disseminated aspergillosis confirmed by necropsy. Paradoxically, galactomannan antigen detection in serum was negative. This is the third case of Aspergillus endocarditis with positive blood culture reported in the literature.

Topics: Amaurosis Fugax; Aneurysm, Infected; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Biomarkers; Caspofungin; Combined Modality Therapy; Echinocandins; Endocarditis; False Negative Reactions; Fatal Outcome; Fungemia; Galactose; Heart Valve Prosthesis Implantation; Humans; Infarction; Kidney; Lipopeptides; Male; Mannans; Mesenteric Arteries; Mesenteric Vascular Occlusion; Middle Aged; Mitral Valve; Peptides, Cyclic; Pulmonary Disease, Chronic Obstructive; Pyrimidines; Renal Artery; Triazoles; Voriconazole

During recent years, a rising incidence of invasive pulmonary aspergillosis (IPA) in non-neutropenic critically ill patients has been reported. Critically ill patients are prone to develop disturbances in immunoregulation during their stay in the ICU, which render them more vulnerable for fungal infections. Risk factors such as chronic obstructive pulmonary disease (COPD), prolonged use of steroids, advanced liver disease, chronic renal replacement therapy, near-drowning and diabetes mellitus have been described. Diagnosis of IPA may be difficult and obtaining histo- or cytopathological demonstration of the fungus in order to meet the gold standard for IPA is not always feasible in these patients. Laboratory markers used as a non-invasive diagnostic tool, such as the galactomannan antigen test (GM), 1,3-beta-glucan, and Aspergillus PCR, show varying results. Antifungal therapy might be considered in patients with persistent pulmonary infection who exhibit risk factors together with positive cultures or sequentially positive GM and Aspergillus PCR in serum, in whom voriconazole is the drug of choice. The benefit of combination antifungal therapy lacks sufficient evidence so far, but this treatment might be considered in patients with breakthrough infections or refractory disease.

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Critical Illness; DNA, Fungal; Drug Therapy, Combination; Galactose; Humans; Intensive Care Units; Lung Diseases, Fungal; Mannans; Opportunistic Infections; Polymerase Chain Reaction; Risk Factors

The usefulness of surrogate markers in the diagnosis of invasive fungal infections caused by Aspergillus and other emerging mycelial fungi is based on the ability of surrogate markers to detect the infection caused by different species of mycelial fungi. Conventional microbiological methods for diagnosis of fungal disease are slow and insensitive. Antigen based assays or measurement of (1-3)-beta-D-glucan in blood have been developed and validated in clinical laboratories. We review these diagnostic contemporary tools, their clinical application and impact.

Topics: Antibodies, Fungal; Antigens, Fungal; Aspergillosis; beta-Glucans; Biomarkers; Clinical Trials as Topic; Communicable Diseases, Emerging; DNA, Fungal; Early Diagnosis; Fungemia; Galactose; Humans; Immunologic Techniques; Mannans; Predictive Value of Tests; Proteoglycans; Risk Factors; Zygomycosis

Invasive aspergillosis is a serious and often lethal fungal infection in immunocompromised patients, with increasing incidence in recent years. The high mortality is related not only to severe immunosuppression but especially to difficulties in early diagnosis. The development of noninvasive nonculture diagnostic methods in recent years is a major advance in the early diagnosis of invasive aspergillosis, but the only method with a clear position is currently galactomannan detection by sandwich ELISA. The test has an excellent negative predictive value and is able to exclude invasive aspergillosis with high probability. In addition, its good sensitivity often allows diagnosis of the condition before it is clinically manifested. However, variations in sensitivity due to numerous factors and potential false-positive results in certain populations are the main limitations to its use. The purpose of this review is to summarize the current knowledge of the use of galactomannan in the early diagnosis of invasive aspergillosis.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Early Diagnosis; Galactose; Humans; Mannans

Pulmonary infiltrates in immunocompromised children often pose problems in terms of deciding on further diagnostic and therapeutic procedures, but few studies have evaluated the value of non-invasive and invasive diagnostic methods in paediatric populations. Both galactomannan ELISA and PCR protocols appear to be less useful in children than in adults. Invasive procedures, such as bronchoalveolar lavage or lung biopsy, can yield a pathohistological diagnosis and/or the isolation of a pathogen. Prospective studies in paediatric patients are needed urgently to assess the value of different diagnostic procedures and to define an effective and safe diagnostic strategy for the individual child.

Topics: Animals; Aspergillosis; Diagnosis, Differential; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Immunocompromised Host; Infant; Lung Diseases; Mannans; Polymerase Chain Reaction

A double-sandwich enzyme-linked immunosorbent galactomannan assay has been approved for surveillance for invasive aspergillosis in immunocompromised patients. We undertook a meta-analysis to assess the accuracy of a galactomannan assay for diagnosing invasive aspergillosis.. Studies of the galactomannan assay that used the European Organization for Research and Treatment of Cancer or similar criteria as a reference standard and provided data to calculate sensitivity and specificity were included. Pooled sensitivity and specificity and summary measures of accuracy, Q* (the upper left-most point on the summary receiver-operating characteristic curve), mean D (a log odds ratio), and Youden index were calculated. Subgroup analyses were performed to explore heterogeneity.. Twenty-seven studies from 1966 to 28 February 2005 were included. Overall, the galactomannan assay had a sensitivity of 0.71 (95% confidence interval [CI], 0.68-0.74) and specificity of 0.89 (95% CI, 0.88-0.90) for proven cases of invasive aspergillosis. The Youden index, mean D, and Q* were 0.54 (95% CI, 0.41-0.65), 2.74 (95% CI, 21.12-3.36), and 0.80 (95% CI, 0.74-0.86), respectively, indicating moderate accuracy. Subgroup analyses showed that the performance of the test differed by patient population and type of reference standard used. Significant heterogeneity was present.. The galactomannan assay has moderate accuracy for diagnosis of invasive aspergillosis in immunocompromised patients. The test is more useful in patients who have hematological malignancy or who have undergone hematopoietic cell transplantation than in solid-organ transplant recipients. Further studies with attention to the impact of antifungal therapy, rigorous assessment of false-positive test results, and assessment of the utility of the test under nonsurveillance conditions are needed.

Topics: Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Patient Selection; Reproducibility of Results

Invasive aspergillosis is extremely rare in immunocompetent children. Here we describe the clinical, radiologic, and laboratory course of fatal invasive pulmonary and central nervous system aspergillosis in a previously healthy child after a near-drowning incident with submersion in a pond. Findings were compared with data from the literature, which is reviewed. Serum Aspergillus galactomannan levels were determined retrospectively and were compared with the results of routine microbiological and radiologic examinations, showing a significant diagnostic and therapeutic delay of the routine diagnostic approach in comparison with the use of the Aspergillus galactomannan assay. This delay may have contributed to the fatal course. Serial determination of serum Aspergillus galactomannan may be helpful in diagnosing invasive aspergillosis early in case of pulmonary disease after near-drowning and may contribute to an early appropriate treatment. Currently voriconazole, eventually in combination with caspofungin, should be considered as the drug of choice in the management of invasive aspergillosis after near-drowning.

Topics: Aspergillosis; Aspergillus fumigatus; Disease Susceptibility; Early Diagnosis; Epilepsy; Fatal Outcome; Female; Fresh Water; Galactose; Gram-Negative Bacterial Infections; Humans; Infant; Lung Diseases, Fungal; Mannans; Near Drowning; Neuroaspergillosis; Paraplegia; Pneumonia, Aspiration; Pneumonia, Bacterial; Respiratory Distress Syndrome; Respiratory Insufficiency; Stenotrophomonas maltophilia; Water Microbiology

Invasive aspergillosis (IA) is a serious cause of morbidity and mortality among immunocompromised patients. Prompt and non-invasive methods for diagnosing IA are needed to improve the management of this life-threatening infection in patients with hematological disorders. In summary, this retrospective review of studies performed on the two assays finds that both assays have high sensitivity and specificity but are more useful when used together as a diagnostic strategy for patients with invasive aspergillosis.

Topics: Aspergillosis; beta-Glucans; Clinical Trials as Topic; Diagnostic Tests, Routine; Galactose; Humans; Lung Diseases, Fungal; Mannans; Multicenter Studies as Topic; Predictive Value of Tests; Proteoglycans; Reproducibility of Results

Invasive aspergillosis (IA) is a leading cause of morbidity and mortality in immunocompromised hosts, with a survival rate lower than 50%. Although proven diagnosis of IA requires histopathological evidence of deep-tissue invasion, eventually supported by microbiological cultures, critical conditions of patients often hamper invasive diagnostic procedures, and many IA cases are not diagnosed during life. Detection of circulating galactomannan antigens is a non-invasive method widely used to support the diagnosis of IA. However, results of test performance have been variable and, in particular, the method shows specificity problems due to cross reactions and false positive results caused by other microorganisms, foods, antibiotics, graft versus host disease and particular physiological conditions of premature infants. It appears therefore mandatory to know exactly all these possible interactions, in order to properly evaluate GM test results and correctly apply them to the diagnosis of IA and to clinical practice.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Cross Reactions; False Positive Reactions; Galactose; Humans; Mannans

Invasive aspergillosis remains an important cause of morbidity and mortality in patients with prolonged and severe immune suppression such as following haematopoietic stem-cell transplantation. Consensus definitions, which allow categorisation of patients based on diagnostic criteria, are an important improvement in uniform registration of invasive mycoses in clinical trials. Prospective monitoring of high-risk patients for the circulating aspergillus cell-wall component galactomannan, results in earlier diagnosis in two-thirds of patients when compared with conventional diagnostic methods. High-resolution CT (HRCT) enables the lesions characteristic of invasive mycoses to be detected earlier and better than by chest radiograph. In addition, invasive mycoses cause characteristic lesions on the HRCT scan including the halo-sign and the air-crescent sign. The pre-emptive management strategy which combines monitoring of patients for surrogate markers with a HRCT scan appears to be a promising approach to the early identification and treatment of patients with invasive aspergillosis.

Topics: Aspergillosis; Biomarkers; Diagnosis, Differential; Galactose; Humans; Immunocompromised Host; Mannans; Risk Factors; Sensitivity and Specificity; Tomography, X-Ray Computed

Within the past decade surrogate markers have become important and reliable tools for the early diagnosis of invasive aspergillosis. Surrogate markers include the antigens galactomannan and (1 --> 3)-beta-D-glucan for which commercial assays are available, as well as fungal DNA for which experimental PCR assays have been developed. Although many clinical validation studies have been performed, the kinetics of these markers are still largely unknown. Recent studies have addressed several issues related to variables that interact with the performance of the assays, including exposure to mould-active antifungal agents and the interpretive cut-off levels. Insights gained as a result of these studies will help us to further optimize management strategies and to determine the optimal test sequence.

Topics: Aspergillosis; Aspergillus; Biomarkers; Clinical Laboratory Techniques; DNA, Fungal; Galactose; Glucans; Humans; Mannans; Polymerase Chain Reaction

Topics: Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillosis, Allergic Bronchopulmonary; Aspergillus; beta-Glucans; Biomarkers; Colorimetry; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Latex Fixation Tests; Lung Diseases, Fungal; Mannans; Nephelometry and Turbidimetry; Polymerase Chain Reaction; Proteoglycans

Invasive aspergillosis occurs in a wide range of clinical scenarios, is protean in its manifestations, and is still associated with an unacceptably high mortality rate. Early diagnosis is critical to a favourable outcome, but is difficult to achieve with current methods. Deep tissue diagnostic specimens are often difficult to obtain from critically ill patients. Newer antifungal agents exhibit differential mould activity, thus increasing the importance of establishing a specific diagnosis of invasive aspergillosis. For these reasons, a range of alternate diagnostic strategies have been investigated. Most investigative efforts have focused on molecular and serological diagnostic techniques. The detection of metabolites produced by Aspergillus spp and a range of aspergillus-specific antibodies represent additional, but relatively underused, diagnostic avenues. The detection of galactomannan has been incorporated into diagnostic criteria for invasive aspergillosis, reflecting an increased understanding of the performance, utility, and limitations of this technique. Measurement of (1,3)-beta-D glucan in blood may be useful as a preliminary screening tool for invasive aspergillosis, despite the fact that this antigen can be detected in a number of other fungi. There have been extensive efforts directed toward the detection of Aspergillus spp DNA, but a lack of technical standardisation and relatively poor understanding of DNA release and kinetics continues to hamper the broad applicability of this technique. This review considers the application, utility, and limitations of the currently available and investigational diagnostic modalities for invasive aspergillosis.

Topics: Antibodies, Fungal; Aspergillosis; Aspergillus; Clinical Laboratory Techniques; DNA, Fungal; Galactose; Humans; Mannans

The availability of the Platelia Aspergillus, a sandwich ELISA kit that detects circulating galactomannan, has been a major advance for managing patients at risk for invasive aspergillosis because of the early detection of the antigen. The assay is now widely used throughout the world, including the USA. Although initial studies that assessed the performance characteristics of this assay reported high sensitivity and specificity, more recent studies show significant variation in performance. The causes of this variability are multifactorial and, in large part, cannot be explained because there is insufficient understanding of the kinetics of galactomannan in vivo. We explored some of the factors that affect the release of the aspergillus antigen that bears the epitope that reacts with the monoclonal antibody used in the ELISA, its leakage from the site of infection into the blood, and its binding to substances present in the blood. Factors that affect the detection of antigen in blood are also discussed, most notably the pretreatment procedure aimed at liberating the antigen from immune complexes. Understanding the biology of galactomannan release by aspergillus will greatly enhance our understanding of the kinetics of this and other surrogate markers and allow their optimum use in the management of invasive aspergillosis.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Reagent Kits, Diagnostic

Invasive aspergillosis is one of the most frequent fungal infections in neutropenic patients, in whom it is associated with a high mortality. Its diagnosis is difficult by the traditionally used laboratory tests. In the last years, an ELISA (Platelia Aspergillus, Bio-Rad, France) to detect galactomannan in neutropenic and cancer patients with high risk of suffering invasive aspergillosis has been developed. The experience accumulated in Spain in the diagnosis of invasive aspergillosis by Platelia Aspergillus is presented in this monograph.

Topics: Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; Fungemia; Galactose; Humans; Immunocompromised Host; Mannans; Neutropenia

Invasive aspergillosis is major cause of morbility and mortality in immunosuppressed patients, in part due to the inability to identify infected patients at an early stage of the disease. Diagnosis is based on a combination of imaging (high-resolution computed tomography) and a number of laboratory techniques including direct examination, culture and circulating markers (galactomannan and Aspergillus DNA) which can be detected at early stages of the infection.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Bone Marrow Transplantation; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Fungemia; Galactose; Humans; Immunocompromised Host; Mannans; Mycology; Neoplasms; Neutropenia; Polymerase Chain Reaction; Sensitivity and Specificity

The usefulness of galactomannan detection using the Platelia Aspergillus test for the diagnosis of invasive aspergillosis was studied in 849 sera from 54 hematological patients with prolonged neutropenia, which were classified according to the risk for invasive aspergillosis. Three patients developed a proven invasive aspergillosis, one a probable invasive aspergillosis and 17 patients a possible invasive aspergillosis. Thirty-three patients showed no evidence of invasive aspergillosis. All patients with proven invasive aspergillosis had a high risk for invasive aspergillosis, while the one having probable invasive aspergillosis had intermediate risk. Detection of galactomannan in this study showed a sensitivity of 66.7% for patients with proven invasive aspergillosis and 50% for patients with proven and probable invasive aspergillosis. The specificity was 98% or higher in all groups studied. The predictive positive and negative values for patients with proven invasive aspergillosis were 66.7% and 98%, respectively. A rise in the concentration of galactomannan was observed in patients who failed to respond to the antifungal treatment. Galactomannan antigenemia preceded post-mortem histological diagnosis of invasive aspergillosis in two patients by 17 and 81 days, respectively. In conclusion, detection of galactomannan by the Platelia Aspergillus test allows for a specific and relatively sensitive diagnosis of invasive aspergillosis in hematological patients with a high and intermediate risk for invasive aspergillosis.

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Enzyme-Linked Immunosorbent Assay; Follow-Up Studies; Fungemia; Galactose; Humans; Immunocompromised Host; Mannans; Neutropenia; Patient Isolators; Predictive Value of Tests; Retrospective Studies; Risk; Sensitivity and Specificity; Time Factors

Invasive aspergillosis (IA) is common in allogeneic SCT recipients, with an incidence of 4-10%. The majority of these infections are diagnosed several months after SCT and they are frequently associated with GVHD. The diagnosis is difficult and often delayed. Established IA is notoriously difficult to treat with a death rate of 80-90%. This review summarises recent data on this problem to assess whether there has been any progress. Effective prophylactic measures are still lacking. Severe immunosuppression is the main obstacle to the success of therapy. Recent and ongoing developments in diagnostic measures and new antifungal agents may improve treatment results to some extent, but Aspergillus infections still remain a formidable problem in allogeneic transplantation. Further studies in this field will focus on the role of various cytokines and combinations of antifungal agents.

Topics: Antifungal Agents; Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; DNA, Fungal; Forecasting; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Incidence; Lung Diseases, Fungal; Mannans; Risk Factors; Sensitivity and Specificity; Transplantation Conditioning; Transplantation, Homologous; Treatment Outcome

Invasive aspergillosis remains a devastating disease, which is partly because of the inability to identify infected patients at an early stage of the disease. Recently, new diagnostic tests and procedures have been developed to help in identifying high-risk patients. High-resolution computed tomography has been shown to be a potent tool for detecting pulmonary abnormalities in neutropenic patients. Assays for the detection of circulating markers such as fungal antigens or Aspergillus DNA have been developed and show that markers can be detected in the blood at an early stage of infection. Also, the markers correlate with the fungal burden in the tissue, which allows monitoring of response to antifungal therapy. The tests and procedures that are now available need to be incorporated into management strategies for at-risk patients and evaluated in clinical trials. Although we now have markers that allow the early detection of fungal products, many questions remain unanswered with respect to the kinetics of the markers in different patient groups, the optimal management strategy and the effect of prophylaxis and treatment on the markers. Nevertheless, the implementation of new approaches for the management of invasive aspergillosis offers opportunities to improve outcome of patients.

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; DNA, Fungal; Galactose; Glucans; Humans; Lung Diseases, Fungal; Mannans; Polymerase Chain Reaction; Sensitivity and Specificity; Tomography, X-Ray Computed

Early diagnosis of invasive aspergillosis is of utmost importance but difficult to achieve. Serological methods were developed mainly because all the other diagnostic approaches had major drawbacks. The detection of antibodies against Aspergillus antigens provided little interesting information, since anti-Aspergillus antibodies were commonly found in healthy people, while, on the other hand, immunocompromised patients often failed to raise antibodies. Several groups of investigators showed that the detection in serum or urine by RIA or ELISA of Aspergillus antigens, was a highly specific indication of invasive disease. The sensitivities of the techniques, however, were moderate, partially due to the low antigen concentrations and the transient character of antigenemia. The only commercially available test is a latex agglutination test detecting 15 ng/ml galactomannan. It has a high specificity but reported sensitivities varied between 42% and 94.7%. Results with an experimental double-sandwich ELISA detecting about 1 ng/ml galactomannan suggest that the improvement of detection limit also greatly increases the clinical value of the test: patients become and remain positive 2 to 10 weeks earlier. If these results are confirmed, antigen detection could become a essential element in the management of an immunocompromised patient population.

Topics: Antibodies, Fungal; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Latex Fixation Tests; Lung Diseases, Fungal; Mannans

Recent studies have shown that the microbiome, namely Aspergillus species, play a previously unrecognized role in both stable and exacerbated chronic obstructive pulmonary disease (COPD). Galactomannan is a major component of the Aspergillus cell wall that has been widely used as a diagnostic marker.. To explore whether serum levels of Aspergillus-galactomannan antigen could be used to evaluate the risk of severe acute exacerbation of COPD (AE-COPD).. We measured the Aspergillus-galactomannan antigen levels of 191 patients with stable COPD, and examined its clinical relevance including AE-COPD.. There were 77 (40.3%) patients who were positive for serum Aspergillus-galactomannan antigen (≥0.5). High Aspergillus-galactomannan antigen level (≥0.7) was associated with older age and presence of bronchiectasis and cysts on computed tomography images. Compared to patients with low Aspergillus-galactomannan antigen level (<0.7), patients with high Aspergillus-galactomannan antigen level had significantly higher incidence of severe AE-COPD (P = 0.0039, Gray's test) and respiratory-related mortality (P = 0.0176, log-rank test). Multivariate analysis showed that high Aspergillus-galactomannan antigen level was independently associated with severe AE-COPD (hazard ratio, 2.162; 95% confidence interval, 1.267-3.692; P = 0.005).. Serum Aspergillus-galactomannan antigen was detected in patients with COPD, and elevated serum Aspergillus-galactomannan antigen was associated with severe AE-COPD.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Aspergillus; Disease Progression; Female; Galactose; Humans; Male; Mannans; Middle Aged; Propensity Score; Pulmonary Disease, Chronic Obstructive; Retrospective Studies; Severity of Illness Index

The benefit of the combination of serum galactomannan (GM) assay and polymerase chain reaction (PCR)-based detection of serum Aspergillus DNA for the early diagnosis and therapy of invasive aspergillosis (IA) in high-risk hematological patients remains unclear.. We performed an open-label, controlled, parallel-group randomized trial in 13 Spanish centers. Adult patients with acute myeloid leukemia and myelodysplastic syndrome on induction therapy or allogeneic hematopoietic stem cell transplant recipients were randomized (1:1 ratio) to 1 of 2 arms: "GM-PCR group" (the results of serial serum GM and PCR assays were provided to treating physicians) and "GM group" (only the results of serum GM were informed). Positivity in either assay prompted thoracic computed tomography scan and initiation of antifungal therapy. No antimold prophylaxis was permitted.. Overall, 219 patients underwent randomization (105 in the GM-PCR group and 114 in the GM group). The cumulative incidence of "proven" or "probable" IA (primary study outcome) was lower in the GM-PCR group (4.2% vs 13.1%; odds ratio, 0.29 [95% confidence interval, .09-.91]). The median interval from the start of monitoring to the diagnosis of IA was lower in the GM-PCR group (13 vs 20 days; P = .022), as well as the use of empirical antifungal therapy (16.7% vs 29.0%; P = .038). Patients in the GM-PCR group had higher proven or probable IA-free survival (P = .027).. A combined monitoring strategy based on serum GM and Aspergillus DNA was associated with an earlier diagnosis and a lower incidence of IA in high-risk hematological patients. Clinical Trials Registration. NCT01742026.

Topics: Adult; Aged; Aspergillosis; Aspergillus; DNA, Fungal; Female; Galactose; Humans; Leukemia, Myeloid, Acute; Male; Mannans; Middle Aged; Myelodysplastic Syndromes; Real-Time Polymerase Chain Reaction; Secondary Prevention

Invasive aspergillosis (IA) is associated with poor outcomes in patients with hematologic malignancies (HMs) and hematopoietic cell transplantation (HCT). Small studies suggest a role for combination antifungal therapy.. To assess the safety and efficacy of voriconazole and anidulafungin compared with voriconazole monotherapy for treatment of IA.. Randomized, double-blind, placebo-controlled multicenter trial. (ClinicalTrials.gov: NCT00531479).. 93 international sites.. 454 patients with HM or HCT and suspected or documented IA were randomly assigned to treatment with voriconazole and anidulafungin or placebo. Primary analysis was done in the modified intention-to-treat population of 277 patients in whom IA was confirmed.. The primary outcome was 6-week mortality; secondary outcomes included 12-week mortality, mortality in major subgroups, and safety measures.. Mortality rates at 6 weeks were 19.3% (26 of 135) for combination therapy and 27.5% (39 of 142) for monotherapy (difference, -8.2 percentage points [95% CI, -19.0 to 1.5]; P  = 0.087). Secondary mortality outcomes favored combination therapy. Multivariable regression analysis suggested that maximum galactomannan value, Karnofsky score, and baseline platelet count had prognostic significance. Most patients (218 of 277 [78.7%]) had IA diagnosis established by radiographic findings and maximum galactomannan positivity. In a post hoc analysis of this dominant subgroup, 6-week mortality was lower in combination therapy than monotherapy (15.7% [17 of 108] vs. 27.3% [30 of 110]; difference, -11.5 percentage points [CI, -22.7 to -0.4]; P = 0.037). Safety measures, including hepatotoxicity, were not different.. Mortality at 6 weeks was higher than expected, and the difference in mortality was lower than expected, which reduced power to detect a treatment effect. Enrollment was restricted to patients with HM or HCT, which limited generalizability.. Compared with voriconazole monotherapy, combination therapy with anidulafungin led to higher survival in subgroups of patients with IA. Limitations in power preclude definitive conclusions about superiority.. Pfizer.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anidulafungin; Antifungal Agents; Aspergillosis; Double-Blind Method; Drug Therapy, Combination; Echinocandins; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Karnofsky Performance Status; Male; Mannans; Middle Aged; Platelet Count; Treatment Outcome; Voriconazole; Young Adult

The utility of Aspergillus galactomannan (GM) and β-D-glucan (BG) in liver transplant recipients remains uncertain.. As part of a randomized, double-blind trial of antifungal prophylaxis in liver transplant recipients at risk for invasive fungal infections (IFIs), GM and BG were assessed in 199 patients at baseline (enrollment) and weekly thereafter for the duration of study drug. Receiver operating characteristic (ROC) analysis was used to evaluate the accuracy of these for the diagnosis of IFIs.. Overall, 46.4% of the patients at baseline had positive GM (index ≥ 0.5) and 89.6% had BG of 80 pg/mL or greater with BG level of 500 pg/mL or greater in 31.8%. Patients with invasive aspergillosis (IA) (3/3) had positive GM at baseline as did 45.5% of those without IA (P = 0.098); the area under the ROC curve for the diagnosis of IA was 0.77 (fair test, ie, good sensitivity but poor specificity). Using BG cutoff of 80 pg/mL or higher, 100% (12/12) of the patients with IFI had positive baseline BG and as did 88.9% (160/180) of those without IFI (P = 0.618); the area under the ROC curve for predicting IFIs was 0.56 (poor test). In multivariate analyses, GM positivity was associated with study site (P = 0.041), and BG positivity with renal replacement therapy (P = 0.05) and study site (P = 0.01). The GM and BG levels declined over time; positivity at subsequent time points was lower in comparison with baseline (P < 0.001).. The GM and BG tests had significant center variability and limited accuracy for the diagnosis of IFIs in high-risk liver transplant recipients.

Topics: Adult; Aged; Antifungal Agents; Aspergillosis; beta-Glucans; Double-Blind Method; Female; Follow-Up Studies; Galactose; Humans; Liver Transplantation; Male; Mannans; Middle Aged; Postoperative Complications; Prospective Studies; Proteoglycans; Risk Factors; ROC Curve; Young Adult

We assessed the success rate of empirical antifungal therapy with itraconazole and evaluated risk factors for predicting the failure of empirical antifungal therapy. A multicenter, prospective, observational study was performed in patients with hematological malignancies who had neutropenic fever and received empirical antifungal therapy with itraconazole at 22 centers. A total of 391 patients who had abnormal findings on chest imaging tests (31.0%) or a positive result of enzyme immunoassay for serum galactomannan (17.6%) showed a 56.5% overall success rate. Positive galactomannan tests before the initiation of the empirical antifungal therapy (P=0.026, hazard ratio [HR], 2.28; 95% confidence interval [CI], 1.10-4.69) and abnormal findings on the chest imaging tests before initiation of the empirical antifungal therapy (P=0.022, HR, 2.03; 95% CI, 1.11-3.71) were significantly associated with poor outcomes for the empirical antifungal therapy. Eight patients (2.0%) had premature discontinuation of itraconazole therapy due to toxicity. It is suggested that positive galactomannan tests and abnormal findings on the chest imaging tests at the time of initiation of the empirical antifungal therapy are risk factors for predicting the failure of the empirical antifungal therapy with itraconazole. (Clinical Trial Registration on National Cancer Institute website, NCT01060462).

Topics: 14-alpha Demethylase Inhibitors; Adolescent; Adult; Aged; Antifungal Agents; Aspergillosis; Candidiasis; Coccidioidomycosis; Febrile Neutropenia; Female; Galactose; Hematologic Neoplasms; Humans; Itraconazole; Male; Mannans; Middle Aged; Prospective Studies; Treatment Outcome; Young Adult

An improved number of anti-fungal drugs are currently available for the treatment of invasive aspergillosis (IA). While serial galactomannan index (GMI) measurement can be used to monitor response to treatment, the extent to which different anti-fungal regimens can affect galactomannan levels is unknown. In 147 IA patients receiving either voriconazole (VCZ) or conventional amphotericin B (CAB) in a multicentre clinical trial, we performed post-hoc analyses of GMI trends in relation to outcomes. The generalized estimation equations approach was used to estimate changes in the effect size for GMI over time within patients. Patients who received VCZ primary therapy and had good treatment response 12 weeks later showed earlier decreases in GMI values at Week 1 and Week 2 (p = 0.001 and 0.046 respectively) as compared to patients who only received CAB. At end-of-randomized therapy (EORT), which was a pre-set secondary assessment point for all patients who switched from randomized primary (CAB or VCZ) to an alternative anti-fungal drug, treatment failure was associated with increasing GMI at Weeks 1 and 2 in CAB-primary treated patients (p = 0.022 and 0.046 respectively). These distinct trends highlight the variations in GMI kinetics with the use of different anti-fungal drugs and their implications in relation to IA treatment response.

Topics: Amphotericin B; Antifungal Agents; Aspergillosis; Biomarkers; Female; Fungal Polysaccharides; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Pyrimidines; Survival Analysis; Treatment Outcome; Triazoles; Voriconazole

Empirical treatment with antifungal drugs is often used in haematology patients at high risk of invasive aspergillosis. We compared a standard diagnostic strategy (culture and histology) with a rapid biomarker-based diagnostic strategy (aspergillus galactomannan and PCR) for directing the use of antifungal treatment in this group of patients.. In this open-label, parallel-group, randomised controlled trial, eligible patients were adults undergoing allogeneic stem-cell transplantation or chemotherapy for acute leukaemia, with no history of invasive fungal disease. Enrolled patients were randomly assigned (1:1) by a computer-generated schedule to follow either a standard diagnostic strategy (based on culture and histology) or a biomarker-based diagnostic strategy (aspergillus galactomannan and PCR) to direct treatment with antifungal drugs. Patients, were followed up for 26 weeks or until death. Masking of the use of different diagnostic tests was not possible for patients, treating physicians, or investigators. The primary endpoint was empirical treatment with antifungal drugs in the 26 weeks after enrolment (for the biomarker-based diagnostic strategy, a single postive galactomannan or PCR result was deemed insufficient to confirm invasive aspergillosis, so treatment in this context was classified as empirical). This outcome was assessed by an independent data review committee from which the study allocations were masked. Analyses were by intention to treat and included all enrolled patients. This study is registered with ClinicalTrial.gov, number NCT00163722.. 240 eligible patients were recruited from six Australian centres between Sept 30, 2005, and Nov 19, 2009. 122 were assigned the standard diagnostic strategy and 118 the biomarker-based diagnostic strategy. 39 patients (32%) in the standard diagnosis group and 18 (15%) in the biomarker diagnosis group received empirical antifungal treatment (difference 17%, 95% CI 4-26; p=0·002). The numbers of patients who had hepatotoxic and nephrotoxic effects did not differ significantly between the standard diagnosis and biomarker diagnosis groups (hepatotoxic effects: 21 [17%] vs 12 [10%], p=0·11; nephrotoxic effects: 52 [43%] vs 60 [51%], p=0·20).. Use of aspergillus galactomannan and PCR to direct treatment reduced use of empirical antifungal treatment. This approach is an effective strategy for the management of invasive aspergillosis in high-risk haematology patients.. Australian National Health and Medical Research Council, Cancer Council New South Wales, Pfizer, Merck, Gilead Sciences.

Topics: Adult; Antifungal Agents; Aspergillosis; Aspergillus; Biomarkers; Female; Follow-Up Studies; Galactose; Humans; Leukemia, Lymphoid; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Stem Cell Transplantation; Transplantation, Homologous

This study was conducted as a prospective, multicenter trial to evaluate the efficacy and safety of micafungin as an empirical therapy for suspected invasive fungal infections (IFIs), including febrile neutropenia (FN), and to evaluate the usefulness of β-D: -glucan (BG) and Aspergillus galactomannan (GM) antigen in patients with hematologic diseases. A total of 121 patients were enrolled and assessed for safety, and 119 were examined for clinical efficacy. The main underlying diseases were acute myeloid leukemia (38.0%), acute lymphoblastic leukemia (18.2%), and malignant lymphoma (18.2%). The median initial daily dose and duration of micafungin treatment were 150 mg/day and 13 days, respectively. The overall response rate for suspected IFIs (n = 119), based on four composite endpoints, including baseline IFI, breakthrough IFIs (proven and probable), survival, and premature discontinuation, was 79.0%. In addition, the response rate for FN (n = 81), based on these four endpoints as well as defervescence during neutropenia, was 39.5%. Breakthrough IFIs (proven, probable, and possible) occurred in five patients during micafungin treatment. All of these patients were positive for either BG or GM before the breakthrough IFIs. The incidence of adverse events (AEs) associated with micafungin was 10.7% and most were mild. The majority of AEs were liver dysfunction. These results indicate the effectiveness and safety of micafungin as an empirical therapy for suspected IFIs, including FN, and the usefulness of monitoring both BG and GM to detect breakthrough IFIs.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antifungal Agents; Antigens, Bacterial; Aspergillosis; beta-Glucans; Candidiasis, Invasive; Chemical and Drug Induced Liver Injury; Early Diagnosis; Echinocandins; Female; Galactose; Hematologic Neoplasms; Humans; Leukemia, Myeloid, Acute; Lipopeptides; Lymphoma; Male; Mannans; Micafungin; Middle Aged; Mycoses; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Young Adult

Invasive fungal infection (IFI) is a serious threat after allogeneic hematopoietic cell transplant (HCT). This multicenter, randomized, double-blind trial compared fluconazole (N = 295) versus voriconazole (N = 305) for the prevention of IFI in the context of a structured fungal screening program. Patients undergoing myeloablative allogeneic HCT were randomized before HCT to receive study drugs for 100 days, or for 180 days in higher-risk patients. Serum galactomannan was assayed twice weekly for 60 days, then at least weekly until day 100. Positive galactomannan or suggestive signs triggered mandatory evaluation for IFI. The primary endpoint was freedom from IFI or death (fungal-free survival; FFS) at 180 days. Despite trends to fewer IFIs (7.3% vs 11.2%; P = .12), Aspergillus infections (9 vs 17; P = .09), and less frequent empiric antifungal therapy (24.1% vs 30.2%, P = .11) with voriconazole, FFS rates (75% vs 78%; P = .49) at 180 days were similar with fluconazole and voriconazole, respectively. Relapse-free and overall survival and the incidence of severe adverse events were also similar. This study demonstrates that in the context of intensive monitoring and structured empiric antifungal therapy, 6-month FFS and overall survival did not differ in allogeneic HCT recipients given prophylactic fluconazole or voriconazole. This trial was registered at www.clinicaltrials.gov as NCT00075803.

Topics: Adolescent; Adult; Aged; Antifungal Agents; Aspergillosis; Child; Child, Preschool; Disease-Free Survival; Double-Blind Method; Drug Monitoring; Fluconazole; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Middle Aged; Mycoses; Myeloablative Agonists; Survival Rate; Transplantation, Homologous; Young Adult

We carried out a prospective study on galactomannan enzyme immuno assay (GEI) (Platelia Aspergillus EIA, Bio-Rad) testing for diagnosis of invasive aspergillosis (IA) in serum and broncho-alveolar lavage (BAL) in 200 patients with hematological malignancies and profound neutropenia. The incidence of proven and probable IA was 6% and 5.5%, respectively. In patients with fever or pneumonia, a single-positive GEI test result (galactomannan index >or= 0.5) had excellent specificity (100%). Sensitivity was relatively low (40%) at onset of fever, but increased to 94.7% after 6 days of fever. In patients with infiltrates in chest X-ray or computed tomography scan (n = 48), GEI testing in BAL had a favorable diagnostic accuracy as compared with GEI testing in serum (sensitivity 100% versus 71%). Our findings indicate that antifungal therapy should be started immediately at onset of fever in neutropenic patients with positive GEI tests. In patients with fever refractory to broad-spectrum antibiotics (>or=6 days of fever), the high diagnostic accuracy makes GEI testing a valuable diagnostic tool and questions the common strategy to carry out antifungal treatment irrespective of diagnostic testing in this situation. Our data also show that GEI testing in BAL can be useful for early diagnosis of IA in patients with hematological malignancies and pulmonary infiltrates.

Topics: Adult; Aged; Anti-Bacterial Agents; Antineoplastic Agents; Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; Early Diagnosis; Female; Fever; Galactose; Hematologic Neoplasms; Humans; Immunoenzyme Techniques; Incidence; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Neutropenia; Radiography; Sensitivity and Specificity

Reported sensitivity of the galactomannan enzyme immunoassay as an early diagnostic test for invasive aspergillosis (IA) has been widely variable, ranging from 29% to 100% in earlier clinical studies.. Studies performed to date have analyzed performance using per-patient calculations, limiting their ability to measure the impact of clinical variables that change over time, such as receipt of preventive antifungal therapy. In our study, performance of the test was calculated in per-patient and per-test analyses in a large cohort of patients at high risk for IA from 2 North American centers. A total of 272 serum samples obtained from 46 patients with IA and 3005 serum samples obtained from 269 control patients were analyzed using multiple index cutoff values to define positivity.. Per-patient calculations yielded sensitivities of 43% and 70% using index cutoff values of 1.5 and 0.5, respectively; specificity decreased from 93% with use of the 1.5 index cutoff to 70% with use of the 0.5 index cutoff. Per-test calculations yielded sensitivities of 31% and 59% and specificities of 99% and 92% using index cutoff values of 1.5 and 0.5, respectively. Receipt of mold-active antifungal drugs on the day of testing decreased sensitivity; samples obtained from patients not receiving prophylactic or empirical antifungal drugs yielded a sensitivity of 89% and a specificity of 92% (with use of an index cutoff value of 0.5).. These findings have direct implications for preventive strategies, because the diagnostic utility of the antigen assay is compromised during receipt of prophylactic or empirical antifungal therapies.

Topics: Adolescent; Adult; Aged; Antifungal Agents; Aspergillosis; Aspergillus; Child; False Negative Reactions; Female; Fluconazole; Galactose; Humans; Immunoenzyme Techniques; Itraconazole; Male; Mannans; Middle Aged; Sensitivity and Specificity

We determined the value of galactomannan (GM) detection in computerized tomography (CT)-based broncho-alveolar lavage (BAL) fluid and serum for the diagnosis of invasive pulmonary aspergillosis (IPA) in haemato-oncological patients with neutropenia. CT of the thorax and BAL were performed systematically at predefined clinical indications. GM was determined by sandwich enzyme-linked immunosorbent assay; the clinicians were unaware of the results. Of 160 patients, 17 patients (10.6%) presented with proven, probable or suspected IPA. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of GM detection in CT-based BAL fluid were all 100%. For GM detection in serially sampled serum, the sensitivity was 47%, the specificity 93%, the PPV 73% and the NPV 82%. A non-blinded follow-up study was performed to validate these results. In this study, 22 of 198 patients (11.1%) presented with IPA, and the sensitivity, specificity, PPV and NPV of GM detection in CT-based BAL fluid were 85%, 100%, 100% and 88% respectively. None of BAL fluids obtained after antifungal treatment of 3 d or more were positive. These results indicate that, when CT is used systematically and at an early stage, GM detection in CT-based BAL fluid has a high PPV for diagnosing IPA early in untreated patients.

Topics: Adult; Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; Follow-Up Studies; Galactose; Hematologic Neoplasms; Humans; Lung Diseases, Fungal; Mannans; Predictive Value of Tests; Prospective Studies; Risk; Sensitivity and Specificity; Tomography, X-Ray Computed

Invasive aspergillosis has become the leading cause of death after allogeneic hematopoietic stem cell transplantation. This is partially due to the lack of a prompt diagnosis. Recently the detection of Aspergillus galactomannan antigen by means an ELISA technique in serum has been described. The objective of this study was to validate its usefulness in the allogeneic hematopoietic stem cell transplantation setting.

Topics: Adolescent; Adult; Amphotericin B; Anemia, Aplastic; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Enzyme-Linked Immunosorbent Assay; False Negative Reactions; Female; Fungemia; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Predictive Value of Tests; Prospective Studies; Transplantation Conditioning; Transplantation, Homologous

Serum galactomannan detection is considered to be a useful test for early diagnosis and follow-up of invasive aspergillosis. From February to September 2002, adult patients hospitalized in our Hematology Unit for receiving intensive chemotherapy and/or hematopoietic stem cell transplant were prospectively studied. We analyzed a total of 760 samples obtained from 100 patients. Eleven patients (11%) having a positive result (OD index >1.5 ng/ml) in two consecutive Platelia Aspergillus tests were considered galactomannan-positive cases. On the other hand, 12 patients (12%) were diagnosed of proven or probable invasive aspergillosis. Sensitivity (66.6%), specificity (95.5%), positive predictive value (72.7%) and negative predictive value (96.7%) were comparable to those of larger series. Galactomannan positivity allowed also to anticipate invasive aspergillosis diagnosis (from two to 17 days before radiographic findings and from two to 15 days before mycological culture). Moreover, kinetics of antigenemia could be useful for assessing therapeutic response. Once accepted galactomannan test as a diagnostic criterium for invasive aspergillosis knowing potential causes of false positive results is of paramount importance.

Topics: Adult; Aged; Antigens, Fungal; Antineoplastic Combined Chemotherapy Protocols; Aspergillosis; Aspergillosis, Allergic Bronchopulmonary; Aspergillus; Biomarkers; Combined Modality Therapy; Enzyme-Linked Immunosorbent Assay; Female; Follow-Up Studies; Fungemia; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Neutropenia; Prospective Studies; Sinusitis

The detection of circulating Aspergillus galactomannan antigen is a useful tool for serodiagnosis of aspergillosis. However, the latex agglutination test for the detection of galactomannan is not completely reliable due to it's low sensitivity. The sandwich ELISA was developed to achieve high sensitivity.. The sandwich immunocapture ELISA was evaluated by testing 56 sero-positive and 56 sero-negative samples of circulating galactomannan detected by LA test retrospectively.. Sixty of the samples were positive for galactomannan as measured by sandwich ELISA. Fifteen samples out of 56 samples negative by LA test were positive by ELISA and 4 samples out of 56 samples positive by LA test were negative by ELISA. Among 47 serum samples positive for anti-Aspergillus antibody, 14 samples were positive by ELISA.. In conclusion, galactomannan may be detected in more samples of by the new sandwich ELISA than by LA test.

Topics: Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Galactose; Humans; Latex Fixation Tests; Mannans; Predictive Value of Tests

The double sandwich ELISA detecting Aspergillus galactomannan (GM) was prospectively evaluated for the diagnosis of invasive aspergillosis (IA) in 50 haematological patients at risk for IA. Serum samples were collected once weekly as long as the risk factors persisted. Six patients had proven or probable IA (3 A. fumigatus, 1 A. flavus, 1 A. niger, 1 A. ustus) and the GM titres were parallel to the clinical evolution of IA. Six of nine patients with suspected IA had at least two consecutive serum GM titres above 1 ng/ml and died with increasing titres, whereas the GM-negative patients survived. Positive GM titres did not anticipate the isolation of fungi. Unfortunately, positive GM titres did not anticipate the initiation of antifungal therapy, based on clinical suspicion. Moreover, if a true-positive result was defined as two consecutive positive serum samples, four patients out of 35 without proven, probable or suspected IA were positive. Then, the rate of false-positive results was high (12%) in the range previously reported. Nevertheless, the GM ELISA appears useful to assess IA and to follow the efficacy of antifungal treatment.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Child; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Fatal Outcome; Female; Galactose; Humans; Leukemia; Male; Mannans; Middle Aged; Neutropenia; Prospective Studies

The clinical utility of Pastorex Aspergillus, a commercially available circulating aspergillus galactomannan antigen detection kit was evaluated. In animal models of pulmonary aspergillosis, it was extremely sensitive for detection of not only A. fumigatus antigen but also antigens of A. flavus and A. niger both in serum and urine. In a preliminary study in autopsied or clinically proven cases with aspergillus infections a high positive rate of antigen detection was obtained when it was applied prospectively. A total of 1,373 clinical samples obtained from patients with suspected aspergillus infection were examined. Among 67 patients giving antigen-positive results in their clinical samples including serum, urine, sputum and others, 17 patients proved to have aspergillus infection by histological or cultural studies. Sixteen patients were judged to be suspicious of aspergillus infection. Measurement of serum (1 --> 3) beta-D glucan by G-test in these cases was well correlated to the results of the antigen detection test. However, the serum showed often antigen-negative even when it was examined at the peak of illness. The rapid clearance of the antigen from circulation was considered to be one of the reasons for this phenomenon. In these cases, antigen detection in samples other than serum, such as urine, sputum, and pleural fluid was also useful. In conclusion, when using Pastorex Aspergillus to obtain the early diagnosis of aspergillus infection frequent examination in different types of samples is recommended.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Galactose; Glucans; Humans; Lung Diseases, Fungal; Mannans; Rats; Reagent Kits, Diagnostic; Serologic Tests

(1-->3)-beta-D-Glucan is one of the major structural components of fungi, and it seems that it can be detected by the fractionated (1-->3)-beta-D-glucan-sensitive component from a Limulus lysate, factor G. We evaluated the concentration of (1-->3)-beta-D-glucan by using factor G and other fungal antigens in 24 patients with clinical evidence of mycosis and 36 healthy subjects. The mean concentration of (1-->3)-beta-D-glucan in the plasma of the healthy subjects was found to be 2.7 +/- 1.9 pg/ml (range, < 6.9 pg/ml), and it was found to be substantially higher in all 11 patients with candidemia (mean, 2,207.4 pg/ml; range, 325.4 to 8,449.0 pg/ml). Eight of those 11 patients with candidemia (73%) were positive for the Cand-Tec heat-labile candida antigen and only 3 patients (27%) were positive for mannan antigen. Three patients with invasive pulmonary aspergillosis were positive for galactomannan and had, in addition, high concentrations of (1-->3)-beta-D-glucan (mean, 323.3 pg/ml; range, 27.0 to 894.0 pg/ml). All 10 patients with cryptococcosis (including 2 patients with probable cryptococcosis) were positive for cryptococcal antigen by the Eiken latex test; however, (1-->3)-beta-D-glucan levels were not elevated in these patients (mean, 7.0 pg/ml; range, < 16.5 pg/ml). Our results indicated that (1-->3)-beta-D-glucan levels are elevated in patients with candidiasis and aspergillosis but not in those with cryptococcosis.

Topics: Adult; Antigens, Fungal; Aspergillosis; beta-Glucans; Candidiasis; Cryptococcosis; Female; Fungemia; Galactose; Glucans; Humans; Limulus Test; Male; Mannans; Middle Aged; Serologic Tests

Invasive aspergillosis is a major threat to immunocompromised individuals. Galactomannan (GM) is used as a biomarker for invasive aspergillosis. Investigations recommended in current guidelines include GM testing of bronchoalveolar lavage (BAL) fluids. GM testing of endotracheal aspirate, the sampling of which is less invasive, less resource-intensive and less aerosol-generating, is not validated. We compared the performance of endotracheal aspirate GM as a screening tool to predict BAL fluid GM-positivity in patients with suspected invasive aspergillosis.. Of each patient, a pair of corresponding endotracheal aspirate and BAL fluid samples was tested and compared for GM results. Two sample sets were included. The first consisted of 140 consecutive BAL fluid/endotracheal aspirate pairs obtained from 133 patients. The pairs of the second sample set (n = 38) were selected based on the criterion that the BAL tested positive for GM. All specimens were obtained in a German 2,000 bed tertiary care center.. Among BAL fluid GM-positive samples, endotracheal aspirate GM demonstrated poor specificity (72%) but high sensitivity (92% in predicting BAL fluid GM of ≥ 0.50 and 91% for BAL fluid GM of ≥ 1.00) and an excellent negative predictive value (98%). The use of a marginally elevated cutoff of 0.63 resulted in an improved specificity (72-81%), without loss of sensitivity.. For screening purposes, one might consider testing endotracheal aspirate for GM, which could help avoid unnecessary BAL.

Topics: Aspergillosis; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Humans; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Mannans; Sensitivity and Specificity

Opportunistic fungal infections are an important cause of morbidity and mortality in immunocompromised patients. Invasive aspergillosis (IA) has an important place among these infections with ~ 250.000 cases annually. Reducing the mortality rate due to invasive aspergillosis is possible with early diagnosis and treatment of the disease. Because of the low sensitivity in microscopic examination, the time consuming of culture growth, and the difficulties in distinguishing colonization/infection, serological methods are frequently used in the diagnosis of invasive aspergillosis. The aim of this study was to determine the diagnostic performance of galactomannan and beta glucan tests for the diagnosis of invasive pulmonary aspergillosis (IPA). Sixty patients, followed up with the suspicion of invasive pulmonary aspergillosis in Gazi University Hospital were included in the study. The clinical classification of the patients was made according to the revised European Organization for Research and Treatment of Cancer and the Mycoses Study Group (EORTC/MSG) criteria. A total of 10 patients were classified as probable invasive aspergillosis and 20 patients were classified as possible invasive fungal disease. Demographic data of the patients and various risk factors were recorded. One hundred and thirty serum and nine bronchoalveolar lavage (BAL) fluid samples were studied with Plateliaᵀᴹ Aspergillus Ag (Bio-Rad, France), Dynamiker Aspergillus Galactomannan and Dynamiker Fungus (1-3)-beta-D-Glucan (Dynamiker, China) kits. Sensitivity and specificity values were calculated according to U.S. Food and Drug Administration (FDA) approved Plateliaᵀᴹ Aspergillus Ag test. According to this study, the most important risk factors in the development of IPA were the use of steroids and immunomodulatory drugs. The sensitivity of the galactomannan test in the probable group was 77.8%, the specificity was 96.7%, the sensitivity of the beta glucan test was 61.1%, and the specificity was 92.6%. When these two tests were evaluated together, it was observed that the sensitivity in the probable group increased to 83.3% and the specificity decreased to 89.3%. The combined use of galactomannan and beta glucan tests increases the diagnostic sensitivity. Although the presence of prolonged neutropenia is an important risk factor for IA, the use of steroids and immunomodulatory drugs should be kept in mind in non-neutropenic patients.

Topics: Aspergillosis; beta-Glucans; Bronchoalveolar Lavage Fluid; Humans; Immunomodulating Agents; Invasive Pulmonary Aspergillosis; Mannans; Sensitivity and Specificity

Invasive aspergillosis (IA) affects mainly patients with hematological malignancies, and early diagnosis is crucial for timely treatment. Most diagnoses are based on clinical and mycological criteria, mostly galactomannan (GM) test in serum or bronchoalveolar fluid, which is performed in case of clinical suspicion or as routine screening in patients at high risk who are not receiving anti-mold prophylaxis, for early detection of IA. The aim of this study was to assess in a real-world setting the efficacy of biweekly serum GM screening for the early detection of IA.. A retrospective cohort that included 80 adult patients treated at the Hematology Department, Hadassah Medical Center, 2016-2020, with a diagnosis of IA. Clinical and laboratory data were collected from patients' medical files and the rate of GM-driven, GM-associated, and non-GM-associated IA was calculated.. There were 58 patients with IA. The rate of GM-driven diagnosis was 6.9%, GM-associated diagnosis was 43.1%, and non-GM-associated diagnosis was 56.9%. The GM test as a screening tool had led to IA diagnosis in only 0.2% of screened serums with a number needed to screen in order to find 1 patient with IA of 490.. Clinical suspicion outweighs GM screening as a tool for early diagnosis of IA. Nevertheless, GM has an important role as a diagnostic tool for IA.

Topics: Adult; Aspergillosis; Early Diagnosis; Hematologic Neoplasms; Humans; Retrospective Studies

A rapid and reliable diagnostic test is needed to reduce mortality through early diagnosis of invasive aspergillosis (IA) in patients with hematological malignancies.. To evaluate the efficacy of serum and bronchoalveolar lavage (BAL) Aspergillus galactomannan lateral flow assay (GM-LFA) in IA diagnosis and determine the correlation of GM-LFA with GM enzyme immunoassay (GM-EIA) in patients with hematological malignancies.. In this prospective multicenter study, we used serum and BAL fluid samples from patients with hematological malignancies and suspected IA and performed GM-LFA and GM-EIA. According to the EORTC/MSGERC criteria, patients were grouped as proven (n = 6), probable (n = 22), possible IA (n = 55), or no IA (n = 88). The performance of serum GM-LFA at 0.5 optical density index (ODI) and area under the curve (AUC) were calculated. Spearman's correlation analysis and kappa statistics were performed to determine the agreement between the tests.. GM-LFA showed an AUC of 0.832 in proven/probable IA (sensitivity [SEN], specificity [SPE], negative predictive value [NPV], and diagnostic accuracy were 75%, 100%, 92.6%, and 93.9%, respectively, at a 0.5 ODI) versus that in no IA. A moderate positive correlation was noted between the GM-LFA and GM-EIA scores (p = 0.01). The observed agreement between the tests at 0.5 ODI was almost perfect (p < 0.001). After excluding patients who received mold-active antifungal prophylaxis or treatment, the SEN, SPE, NPV, and diagnostic accuracy for proven/probable IA were 76.2%, 100%, 93.3%, and 94.5%, respectively.. Serum GM-LFA demonstrated high discriminatory power and good diagnostic performance for IA in patients with hematological malignancies.

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Hematologic Neoplasms; Humans; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Mannans; Prospective Studies; Sensitivity and Specificity

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Cell-Free Nucleic Acids; Humans; Mannans; Polymerase Chain Reaction

Invasive aspergillosis (IA) in immunocompromised hosts carries high morbidity and mortality. Diagnosis is often delayed because definitive diagnosis requires invasive specimen collection, while noninvasive testing with galactomannan is moderately accurate. Plasma cell-free DNA polymerase chain reaction (cfDNA PCR) represents a novel testing modality for the noninvasive diagnosis of invasive fungal disease (IFD). We directly compared the performance of Aspergillus plasma cfDNA PCR with serum galactomannan for the diagnosis of IA during routine clinical practice.. We conducted a retrospective study of all patients with suspected IFD who had Aspergillus plasma cfDNA PCR testing at Stanford Health Care from 1 September 2020 to 30 October 2022. Patients were categorized into proven, probable, possible, and no IA based on the EORTC/MSG definitions. Primary outcomes included the clinical sensitivity and specificity for Aspergillus plasma cfDNA PCR and galactomannan.. Overall, 238 unique patients with Aspergillus plasma cfDNA PCR test results, including 63 positives and 175 nonconsecutive negatives, were included in this study. The majority were immunosuppressed (89.9%) with 22.3% 30-day all-cause mortality. The overall sensitivity and specificity of Aspergillus plasma cfDNA PCR were 86.0% (37 of 43; 95% confidence interval [CI], 72.7-95.7) and 93.1% (121 of 130; 95% CI, 87.4-96.3), respectively. The sensitivity and specificity of serum galactomannan in hematologic malignancies/stem cell transplants were 67.9% (19 of 28; 95% CI, 49.3-82.1) and 89.8% (53 of 59; 95% CI, 79.5-95.3), respectively. The sensitivity of cfDNA PCR was 93.0% (40 of 43; 95% CI, 80.9-98.5) in patients with a new diagnosis of IA.. Aspergillus plasma cfDNA PCR represents a more sensitive alternative to serum galactomannan for noninvasive diagnosis of IA.

Topics: Aspergillosis; Aspergillus; Cell-Free Nucleic Acids; Humans; Invasive Fungal Infections; Mannans; Polymerase Chain Reaction; Retrospective Studies; Sensitivity and Specificity

Topics: Aspergillosis; Child; Galactose; Humans; Leukemia, Myeloid, Acute; Mannans

This study aims to determine the diagnostic utility of galactomannan enzyme immunoassay (GM EIA) in invasive aspergillosis (IA) in children with hematological malignancy (high risk population) in terms of sensitivity, specificity, negative predictive value (NPV) and positive predictive values (PPV) at various cut offs while validating the revised EORTC/MSG 2019 criteria in order to obtain the best cut-off.. For 100 pediatric patients, serum and respiratory samples were collected. Clinical, mycological workup (potassium-hydroxide mount, fungal culture) and GM EIA was done to classify proven, probable, and possible IA as per EORTC-MSG guidelines,2019. Sensitivity, specificity, PPV and NPV were calculated of GM indices at cut-off 0.5, 0.7 and 1, and validated with revised EORTC -MSG, 2019.. Of 100 patients enrolled, 75 were diagnosed with ALL, 14 with AML, two with Hodgkin's, three had non-Hodgkin lymphoma, and six had undifferentiated leukemia. With routine mycological findings, 51 were classified as probable IA, 11 as possible IA, and 38 as no IA. Aspergillus flavus was the most prevalent on culture (56.9%, 29/51) followed by A. fumigatus (29%, 15/51) A. niger (7.8%, 4/51), A. terreus (3.9%, 2/51) and A. nidulans (2%, 1/51). GM EIA demonstrated sensitivity 82.3%, specificity 97.4%, PPV 98.1%, and NPV 77.1% at cut-off 0.67 when comparing probable/possible IA v/s no IA groups. The GM EIA had the best sensitivity (82.4%), specificity (81.8%), PPV (95.5%), and NPV (50%) at cut off 0.78 when the probable IA group was compared to the possible IA. Seven patients succumbed of whom 5 had GMI ≥ 2.. This study deduces the optimal cut-off for serum GM EIA to be 0.67 obtained by ROC analysis when comparing possible and probable IA versus no IA and reinforces the definition of probable category of EORTC-MSG criteria, 2019. At 0.5 ODI the sensitivity (87.1%) and NPV (80.5%) are high, thus making it the most suitable cut-off for detecting true positive and ruling out IA respectively, in pediatric patients with hematological malignancy. GM EIA when performed adjunctive to clinico-radiological findings can prove to be screening, diagnostic and prognostic test for IA in pediatric hematological malignancy patients.

Topics: Aspergillosis; Child; Hematologic Neoplasms; Humans; Immunoenzyme Techniques; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Mannans; Sensitivity and Specificity

Aspergillosis is an opportunistic systemic infection caused by members of. A case-control study was performed to involve 184 subjects, distributing in four groups: 55 patients with RA, 54 with asthma, 27 with both RA and asthma, and 48 healthy individuals. Serum was collected from involved subjects for detection of human. Aspergillosis was more frequently diagnosed in females with RA and both RA and asthma in opposite to the males. It also was found in most common in middle-aged subjects. There was no significant difference in measurement of GM between all patient groups and healthy individuals.. Aspergillosis can develop in either immunocompetent or immunocompromised individuals. Patients with either RA or RA and asthma are more susceptible to acquired aspergillosis than those with only one disease. Application of GM for diagnosis of aspergillosis may show a nonsignificant results when it uses alone and needs other investigation tests.

Topics: Arthritis, Rheumatoid; Aspergillosis; Aspergillus; Asthma; Case-Control Studies; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Sensitivity and Specificity

Invasive aspergillosis (IA) is an important cause of morbidity and mortality in immunosuppressed children. Early detection of the infection can improve prognosis in this patient population.. To investigate the utility of. For the study, 659 GM-EIA results from 59 patients diagnosed with IA and 3368 GM-EIA results from 351 subjects without evidence for IA (controls) were reviewed retrospectively. Three cut-off values (i.e. ≥0.5, ≥1, ≥1.5) were specified to determine GM-EIA positivity.. The GM-EIA may be used for both screening and diagnostic purposes in paediatric patients using a cut-off value of ≥1.5 for GM-EIA positivity.

Topics: Aspergillosis; Child; Female; Galactose; Humans; Invasive Fungal Infections; Male; Mannans; Retrospective Studies; Sensitivity and Specificity

The use of nonculture-based biomarkers such as the determination of galactomannan is sought for the diagnosis of invasive aspergillosis. To investigate the comparative yield of two tests for the detection of galactomannan in patients with or without proven or probable invasive aspergillosis. Overall, 327 samples (327 patients) were analyzed in a retrospective/prospective study performed in 3 hospitals in Madrid, comparing the determination results in serum or bronchoalveolar lavage of two techniques for galactomannan detection, namely, Platelia Aspergillus Ag (Bio-Rad) and Aspergillus galactomannan Ag Virclia Monotest (Vircell S.L.), following the manufacturer's instructions. Both techniques can automate the process, but the second technique has the advantage of individual processing and assembly of each sample without the need for the additional expense of single-dose strips in controls. In total, 288 of the 327 tests performed showed concordant results between both techniques. The agreement between both methods was к = 0.722, and the correlation between indices was ρ = 0.718. Only 39 samples showed discordant results. In those 39 cases, there were 15 patients with proven or probable invasive aspergillosis criteria. For the samples with clinical criteria as a reference, the areas under the curve of the receiver operating characteristic (ROC) curve were 0.962 for Platelia and 0.968 for VirClia. The VirClia test has been proven to be an alternative for diagnosis due to its friendlier automated format than that of the usual Platelia routine test. The VirClia test also allows individual action and, therefore, a more immediate clinical response.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Humans; Mannans; Prospective Studies; Retrospective Studies; Sensitivity and Specificity

The incidence of Aspergillus and Candida CNS infection, which are characterised by high mortality rates, is underestimated. This underdiagnosis presumably results from the limitations of available diagnostic tools and the need for invasive sampling. Little is known about the role of serologic biomarkers in the setting of CNS aspergillosis and candidiasis.. Serum and cerebrospinal fluid (CSF; 10) samples of 19 patients, whose CNS specimens yielded growth of Aspergillus or Candida, were analysed for different biomarkers for fungal infection, that is galactomannan (GM), galactomannoprotein (GP), mannan, anti-mannan-antibodies and β-1,3-D-glucan (BDG). Serum and CSF specimens of time-matched patients (two each for every case of fungal CNS infection) were included as controls.. Galactomannan, GP and BDG seropositivity was found in one, two and three of five cases of CNS aspergillosis. BDG and mannan/anti-mannan-antibody sensitivity in proven CNS candidiasis was 40% and 20%, respectively. Applying the serum cut-off, sensitivity in CSF testing was 100% for GM and BDG and 50% for mannans. While serum specificity for all assays ranged from 89 to 97%, specificity for CSF BDG was only 70%. No false-positive GM results from CSF were obtained.. Sensitivity for diagnosing CNS aspergillosis and CNS candidiasis from serum is mediocre for all serological biomarkers. GM testing in CSF proved excellent performance. With a sensitivity of 100% but a specificity of only 70%, CSF BDG might be most useful when used in patients with a high pre-test probability.

Topics: Aspergillosis; Aspergillus; beta-Glucans; Biomarkers; Candida; Candidiasis; Central Nervous System; Galactose; Humans; Mannans; Sensitivity and Specificity

To compare a lateral-flow device (LFD) method to the galactomannan assay (GM) for the diagnosis of invasive aspergillosis (IA).. First, 20 GM-positive serum samples stored for two years were retested with both the GM and LFD assays. Second, 153 serum samples from 91 immunocompromised patients suspected of having IA were tested prospectively, including 56 hematologic malignancies and 35 chronic illnesses with steroid therapy.. For the twenty GM-positive stored samples, only ten were positive for the repeated GM assay and none were positive for IA according to the LFD test. The concordance of the LDF with the GM test was 79.81% (83/104) if both tests were performed on the sample collection day, with the rate reducing to 67.65% (23/34) (p < 0.05) if the LFD test was performed 2-7 days after the GM test. Furthermore, there was a significant difference in the discrepancy between the GM and LFD tests between previous and no anti-mold exposure subgroups (33.33% vs. 12.31%, p < 0.01). The sensitivity and specificity of the GM test were 89.65% and 98.66%, 68.96%, and 78.67% for the LFD assay.. Serum samples that have been stored long term are not suitable for re-testing with the GM or LFD assay. There was a strong correlation between the LFD and GM assay results if the tests were performed on the same day, however, this decreased if the samples were stored for more than 2 days. Additionally, previous exposure to antibiotics and/or antifungal therapy could influence the LFD results, leading to discrepancies with the GM test results.

Topics: Anti-Bacterial Agents; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Humans; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Mannans; Sensitivity and Specificity; Steroids

Early diagnosis of invasive aspergillosis is an important factor to improve survival but remains challenging. The detection of Aspergillus antigens is included in the consensus case definitions of the European Organization for Research and Treatment of Cancer and the National Institute of Allergy and Infectious Diseases Mycoses Study Group as a criterion of "probable" invasive aspergillosis. JF5, a mouse IgG3 monoclonal antibody detecting an Aspergillus mannoprotein, has already been implemented as a lateral flow device (LFD). Now, also a JF5-based enzyme-linked immunosorbent assay (EIA) is commercialized (Aspergillus specific galactomannoprotein [GP] EIA, Euroimmun Medizinische Labordiagnostika AG). In this study, we analyzed the diagnostic performance of GP in 63 bronchoalveolar lavage fluid (BALf) samples and 224 serum samples and compared it to performance of the galactomannan (GM) (Platelia Aspergillus enzyme immunoassay (EIA) (Bio-Rad, Marnes-la-Coquette, France)) and the JF5-based LFD (AspLFD; OLM Diagnostics, Newcastle Upon Tyne, United Kingdom). The diagnostic performance of GP and GM correlated well with both having high specificity. With an optimized cutoff threshold for positivity of 0.4-deviating from the 0.5 threshold recommended by the manufacturer-sensitivity of GP in serum is not significantly different than that of GM. However, in BALf sensitivity of GP is significantly less than for GM.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Enzyme-Linked Immunosorbent Assay; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Mannans; Mice; Sensitivity and Specificity

The use of galactomannan (GM) testing in plasma and bronchoalveolar lavage fluid (BALF) has improved the diagnosis of invasive pulmonary aspergillosis (IPA) in patients with chronic obstructive pulmonary disease (COPD); however, the high false-positive rate leads to overdiagnosis. Pentraxin 3 (PTX3) as an indicator of inflammation plays an important role in resistance to Aspergillus infections. This study aimed to investigate the diagnostic value of PTX3 for diagnosing IPA with COPD.. We retrospectively collected data on patients with suspected COPD and IPA who had been hospitalized in the Third Affiliated Hospital of Soochow University between September 2017 and November 2020. PTX3 and GM were measured using enzyme-linked immunosorbent assays.. A total of 165 patients were included in the study, of whom 35 had confirmed or probable IPA. The remaining 130 patients served as controls. The median plasma and BALF PTX3 levels were significantly higher in patients with IPA than in control patients (3.74 ng/mL vs. 1.29 ng/mL, P < 0.001; and 3.88 ng/mL vs. 1.58 ng/mL, P < 0.001 in plasma and BALF, respectively). The plasma GM, plasma PTX3, BALF GM, and BALF PTX3 assays had sensitivities of 60.0%, 77.1%, 78.6%, and 89.3%, respectively, and specificities of 73.8%, 69.2%, 80.7%, and 77.1%, respectively. The sensitivity of PTX3 in plasma and BALF was higher than that of GM. However, the specificity of PTX3 and GM did not differ significantly between the IPA group and the control group. The specificity of the assays for the diagnosis of IPA was > 90% in patients who were PTX3-positive and GM-positive in plasma and BALF.. BALF and plasma PTX3 levels were significantly higher in COPD patients with IPA. The sensitivity of PTX3 was superior to that of GM for diagnosing IPA in patients with COPD. The combination of GM and PTX3 is useful for the diagnosis of IPA in patients with COPD.

Topics: Aged; Aged, 80 and over; Aspergillosis; Biomarkers; Bronchoalveolar Lavage Fluid; C-Reactive Protein; China; Diagnosis, Differential; Female; Galactose; Humans; Male; Mannans; Middle Aged; Pulmonary Disease, Chronic Obstructive; Retrospective Studies; Serum Amyloid P-Component

Aspergillus species meet the most important group of invasive fungal diseases (IFD) in immunosuppressed patients. Galactomannan is a polysaccharide antigen located in the wall structure of Aspergillus. The most commonly used method for antigen detection is enzyme-linked immunoassay (ELISA). Aspergillus galactomannan lateral flow assay (LFA) constitutes one of the new methods in the diagnosis of invasive aspergillosis (IA). The goal of this study was to demonstrate efficacy of LFA in our patients and to compare it to synchronous ELISA results.. Galactomannan antigen was examined using both LFA and ELISA in serum samples taken from patients who were followed up in our haematology clinic. All patients are classified in subgroups as 'proven', 'probable' and 'possible' patients according to the last EORTC / MSG guideline. Patients who met the 'proven' IA criteria were included in the study as the gold standard.. A total of 87 patients were included in the study. Majority of patients had acute myeloid leukaemia (AML) (56.3%). Eleven (12.6%) were in 'proven' IA group. LFA test showed a superior diagnostic performance compared with ELISA (LFA. The most important finding of this study is that the specificity of LFA was found to be higher for cut-off value of 0.5. It is recommended to combine the methods in many studies to provide a better early diagnosis for IA.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Diagnostic Tests, Routine; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Immunocompromised Host; Invasive Fungal Infections; Leukemia, Myeloid; Male; Mannans; Middle Aged; Sensitivity and Specificity

Aspergillus infection is a well-known complication of severe influenza and severe acute respiratory syndrome coronavirus (SARS-CoV), and these infections have been related with significant morbidity and mortality even when appropriately diagnosed and treated. Recent studies have indicated that SARS-CoV-2 might increase the risk of invasive pulmonary aspergillosis (IPA). Here, we report the first case of Aspergillus ochraceus in a SARS-CoV-2 positive immunocompetent patient, which is complicated by pulmonary and brain infections. Proven IPA is supported by the positive Galactomannan test, culture-positive, and histopathological evidence. The patient did not respond to voriconazole, and liposomal amphotericin B was added to his anti-fungal regimen. Further studies are needed to evaluate the prevalence of IPA in immunocompetent patients infected with SARS-CoV-2. Consequently, testing for the incidence of Aspergillus species in lower respiratory secretions and Galactomannan test of COVID-19 patients with appropriate therapy and targeted anti-fungal therapy based on the primary clinical suspicion of IPA are highly recommended.

Topics: Adult; Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus ochraceus; Biomarkers; Brain Abscess; Bronchoalveolar Lavage Fluid; COVID-19; COVID-19 Nucleic Acid Testing; Fatal Outcome; Galactose; Humans; Immunocompetence; Invasive Fungal Infections; Lung Diseases, Fungal; Male; Mannans; SARS-CoV-2; Voriconazole

The consensus definitions of invasive fungal diseases from the EORTC/MSGERC were recently revised and updated. They now include consensus cutoff values for the galactomannan test that support the diagnosis of probable invasive aspergillosis. In this supplement article, we provide a rationale for these proposed thresholds based on the test's characteristics and performance in different patient populations and in different specimen types.

Topics: Antigens, Fungal; Aspergillosis; Consensus; Galactose; Humans; Invasive Fungal Infections; Mannans; Sensitivity and Specificity

To explore the diagnostic value of a galactomannan (GM) detection for non-immunocompromised critically ill patients with influenza-associated aspergillosis (IAA). In this retrospective case-control study, we explored the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and area under the receiver operating characteristic (ROC) curve (AUC) of serum and bronchoalveolar lavage fluid (BALF) GM tests by four detection strategies at different detection time points and with different compound modes. In total, 90 patients were evaluated. The AUC values of the second serum GM test, the first and second BALF GM tests, were significantly higher (0.839 (95% CI 0.716 to 0.963), P < 0.01; 0.904 (95% CI 0.820 to 0.988), P < 0.01; 0.827 (95% CI 0.694 to 0.961), P = 0.043) than that of the first serum GM test (0.548 (95% CI 0.377 to 0.718)). We found that at least one positive result on two consecutive serum GM tests (0.719 (95% CI 0.588 to 0.849)) was the best compared with the first positive test (0.419 (95% CI 0.342 to 0.641), P < 0.01) and positives on two consecutive tests (0.636 (95% CI 0.483 to 0.790), P = 0.014). However, there were no differences between those three detection strategies of BALF GM. The BALF GM test might have a better diagnostic value for IAA in the ICU than the serum GM test. A possible cutoff value of 1.0 to 1.3 was set for GM from BALF specimens for IAA. A single serum GM test is not routinely recommended, but at least one positive result on two consecutive tests appeared to be useful.

Topics: Adult; Aged; Aspergillosis; Bronchoalveolar Lavage Fluid; Case-Control Studies; Clinical Laboratory Techniques; Critical Illness; Female; Galactose; Humans; Influenza, Human; Invasive Pulmonary Aspergillosis; Male; Mannans; Middle Aged; Predictive Value of Tests; Retrospective Studies; ROC Curve; Seasons; Sensitivity and Specificity

Establishing the diagnosis of invasive mold infections (IMI) in immunocompromised children is challenging due to nonspecific clinical presentations and the limited sensitivity of traditional culture-based methods. Rapid non-culture-based diagnostics such as the 1,3-beta-d-glucan and galactomannan assays have emerged as promising adjuncts to conventional diagnostic tests in adults. Available data suggest that 1,3-beta-d-glucan has limited accuracy in the pediatric population and is not recommended to be used for the diagnosis of IMI in children. On the other hand, the diagnostic performance of the serum and bronchoalveolar lavage galactomannan in immunocompromised children is comparable to results observed in adults and can be used as a screening tool in children at high risk of developing invasive aspergillosis (IA) who are not receiving mold-active antifungal prophylaxis and as a diagnostic tool in symptomatic children suspected of having IA. Herein, we summarize the available evidence for the use of these rapid non-culture-based diagnostics in immunocompromised children. We also summarize potential causes of false positivity for the 1,3-beta-d-glucan and galactomannan assays.

Topics: Adult; Aspergillosis; beta-Glucans; Child; Galactose; Glucans; Humans; Immunocompromised Host; Mannans; Sensitivity and Specificity

Topics: Antifungal Agents; Aspergillosis; Galactose; Humans; Mannans; Voriconazole

The aim of this study was to evaluate the diagnostic utility of serum galactomannan (GM) positivity for invasive aspergillosis (IA) in children. Positive GM results between January 2015 and August 2017 were reviewed retrospectively in children with hematologic malignancies. Single and consecutive positive GM results were evaluated according to the different galactomannan index (GMI) (>0.5, >0.7, >1.0 and >1.5) values. There were 104 positive GM results of 70 patients. IA was identified in 29 patients (41.4%) (2 proven and 27 probable). For a single positive GMI of >0.5, >0.7, >1.0, and >1.5, the numbers were 104, 76, 57, and 32 and the positive predictive values (PPVs) were 39.4%, 43.2%, 47.2%, and 50.0%, respectively. The single GM positivity at different thresholds showed no difference between the IA and non-IA group (P>0.05). For 2 consecutive positive GMI values of >0.5, >0.7, >1.0, and >1.5, the numbers were 34, 20, 13, and 4, and the PPVs were 58.8%, 65.0%, 84.6%, and 100.0%, respectively. In the IA group, positivity was higher at all thresholds (P<0.05). According to our findings, consecutive GM positivity has higher PPVs independently from the cutoff value chosen. In pediatric patients with high risk, consecutive sampling should be preferred.

Topics: Adolescent; Aspergillosis; Aspergillus; Biomarkers; Child; Child, Preschool; Cross-Sectional Studies; Female; Follow-Up Studies; Galactose; Hematologic Neoplasms; Humans; Infant; Male; Mannans; Prognosis; Retrospective Studies; Tertiary Care Centers; Turkey

The introduction of non-culture-based diagnostic techniques is revolutionizing the world of microbiological diagnosis and infection assessment. Fungi are no exception, and the introduction of biomarkers has opened up enormous expectations for better management of these entities. Biomarkers are diverse, their targets are also diverse and their evaluation has been done preferably in an individualized use and with deficient designs. Less is known about the value of the combined use of biomarkers and the impact of the negativity of two or more biomarkers on antifungal treatment decisions has been poorly studied. Given the paucity of prospective, randomized and definitive studies, we have convened experts from different fields, with an interest in invasive fungal infections, to answer some questions about the current relevant use of fungal biomarkers. This document summarizes the answers of these experts to the different questions.

Topics: Antibodies, Fungal; Aspergillosis; Aspergillus; Biomarkers; Bronchoalveolar Lavage; Candida; Candidemia; False Positive Reactions; Fluorescent Antibody Technique, Indirect; Galactose; Glucans; Humans; Intensive Care Units; Invasive Fungal Infections; Mannans; Sensitivity and Specificity; Spain

Methodologies to identify epitopes or ligands of the fungal cell wall polysaccharides influencing the immune response of human pathogens have to date been imperfect. Using the galactomannan (GM) of

Topics: Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Cell Wall; Epitopes; Galactose; Humans; Immunomodulation; Mannans; Oligosaccharides

Central nervous system aspergillosis is relatively rare and difficult to diagnose. Here, we report a case of 90-year-old man with chronic lymphocytic leukemia who presented with a month-long gradually worsening headache followed by 3 days of low-grade fever associated with altered mental status. Aspergillus meningitis diagnosed using Aspergillus galactomannan antigen in the cerebrospinal fluid and treated with voriconazole. Delayed diagnosis and treatment of Aspergillus meningitis is typically associated with high mortality; therefore, it is imperative to include this disease in the differential diagnoses of subacute meningitis.

Topics: Aged, 80 and over; Antifungal Agents; Antigens, Fungal; Aspergillosis; Diagnosis, Differential; Galactose; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Male; Mannans; Meningitis, Fungal; Voriconazole

To assess the utility of galactomannan and beta-D-glucan assays in the diagnosis of invasive aspergillosis in clinically suspected cases, and to compare their diagnostic potential to determine whether a combination of the two may result in an early and specific diagnosis.. The descriptive cross-sectional case-control study was conducted at the Armed Forces Institute of Pathology, Rawalpindi, Pakistan, from April 1, 2017, to March 31, 2018, and comprised serum samples from clinically suspected invasive aspergillosis patients and healthy controls. The sera were tested for galactomannan and beta-D-glucan detection. Proven, probable and possible categories of invasive aspergillosis according to European Organisation for Research and Treatment of Cancer / Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group criteria. Galactomannan antigen was detected using a one-stage immunoenzymatic sandwich microplate assay. Beta-D-Glucan antigen was detected using a protease zymogen-based colorimetric assay. Sensitivity and positive / negative likelihood ratio of both the cases and the controls were calculated and compared.. Of the 178 subjects, 119(67%) were cases and 59(33%) were controls. Beta-D-glucan assay was more sensitive than galactomannan assay (91.6% versus 80.67%) whereas galactomannan assay was more specific than beta-D-glucan assay (86.44% versus 76.27%) in the diagnosis of invasive aspergillosis. The sensitivities of both assays decreased with decreasing probability of invasive aspergillosis, i.e., maximum sensitivities of both beta-D-glucan and galactomannan assays were for proven cases (100% versus 87.5%), followed by probable cases (89.29% versus 85.71%), and possible cases (91.57% versus 78.31%).. Both beta-D-glucan and galactomannan assays seemed to play an encouraging role in the diagnosis of invasive aspergillosis in high-risk clinically suspected cases, with the former assay being more sensitive and the latter assay being more specific.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Early Diagnosis; Galactose; Humans; Immunoenzyme Techniques; Invasive Fungal Infections; Mannans; Sensitivity and Specificity

Eye damage during invasive aspergillosis is rarely described and biological diagnosis remains challenging. Here we report the case of a heart transplant recipient with ocular aspergillosis complicating disseminated aspergillosis. Although voriconazole was rapidly given, a decrease in visual acuity of the right eye was consistent with endophthalmitis, resulting in an emergency vitrectomy. The diagnosis was rapidly confirmed: laboratory results showed the presence of Aspergillus fumigatus in a vitreous sample. A series of systemic antifungal medications (liposomal amphotericin B, caspofungin, and voriconazole), several liposomal amphotericin B ocular injections, and pars plana vitrectomy resulted in a limited positive clinical outcome. Interestingly although standard mycological follow-up procedures were negative, Aspergillus antigen testing gave an index of 5.92 on vitreous humour, thus a new intraocular injection of liposomal amphotericin B was performed and voriconazole reinitiated. Ten other vitreous samples from patients without fungal infections were also tested, all showing indexes below 0.25. Although larger studies are needed, this case illustrates that galactomannan testing of vitreous humour could be useful for the diagnosis of fungal endophthalmitis if these data are confirmed in other patients, in particular, if standard mycology is negative and PCR is not available.

Topics: Adult; Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Endophthalmitis; Eye Infections, Fungal; Female; Galactose; Humans; Male; Mannans; Middle Aged; Visual Acuity; Vitrectomy; Vitreous Body; Voriconazole

Invasive fungal disease is a major cause of morbidity and mortality in children with cancer and high-risk febrile neutropenia (HRFN). Repeated serum galactomannan (sGM) measurements have been described as an effective tool to guide therapy in adults under suspicion of invasive aspergillosis. However, the utility of this approach has not been reported in paediatric population.. To evaluate the usefulness of sGM measurements in initiating and modifying antifungal therapy (AFT) in children with cancer and persistent HRFN.. Nested case-control study in children with cancer and persistent HRFN episodes, between July 2013 and January 2019. Patients were classified as cases and controls depending on if they received AFT or not, respectively. Through odds ratio analysis, we assessed the role of sGM positivity in the AFT initiation decision. Then, we analysed the group of patients that initiated AFT, and compared those who had AFT modifications and those who did not, analysing different sGM kinetics thresholds.. A total of 191 episodes from children with persistent HRFN were enrolled, of which 107 received AFT and 84 did not. The median age was 7 years (IQR 4-12), 52% were male and 89% had a haematologic malignancy as underlying disease. Positive sGM was not associated with AFT initiation (OR 0.99, 95% CI 0.43-2.33, P = .99). A difference threshold in sGM Δ ≥ 0.3 sGM was significantly associated with AFT modification (OR 5.07, 95% CI 1.02- 25.70, P = .04).. Our results suggest the utility of serial sGM sampling during AFT in children with persistent HRFN.

Topics: Antifungal Agents; Aspergillosis; Case-Control Studies; Chemotherapy-Induced Febrile Neutropenia; Child; Female; Galactose; Hematologic Neoplasms; Humans; Invasive Fungal Infections; Invasive Pulmonary Aspergillosis; Male; Mannans; Neoplasms

Even though both cellular and humoral immunities contribute to host defense, the role played by humoral immunity against the airborne opportunistic fungal pathogen

Topics: Aspergillosis; Aspergillus fumigatus; beta-Glucans; Bronchoalveolar Lavage Fluid; Cell Wall; Complement Activation; Complement C3; Cytokines; Fungal Polysaccharides; Galactose; Host Microbial Interactions; Humans; Immunity, Cellular; Immunity, Humoral; Integrin alphaXbeta2; Macrophage-1 Antigen; Macrophages; Mannans; Opsonin Proteins; Phagocytosis; Primary Cell Culture; Protein Binding; Reactive Oxygen Species; Serum; Spores, Fungal

The incidence of invasive aspergillosis (IA) has dramatically increased during the last decade. This infection is associated with high morbidity and mortality, ranging from 30% to 70%, especially in immunocompromised patients. Delay in diagnosis and treatment is usually associated with high mortality rates. This study was aimed to assess the diagnostic value of Galactomannan EIA (GM) for early diagnosis of aspergillosis in hospitalized patients with underlying conditions. Also, the antifungal drug susceptibility profiles of causative agents were investigated. In this descriptive cross-sectional study, during the period of 18 months starting from September 2017 until February 2019, 22 bronchoalveolar lavage (BAL) and 13 biopsies from infected sinuses were obtained from a total of 150 patients suffering from different types of hematologic malignancies. All the samples were subjected to microscopic examination and fungal culture. Also, serum specimens were obtained from all patients (n = 135). 22 serum and 17 BAL specimens were tested for the GM level. Fungal identified were confirmed through the PCR-sequencing of the β-tubulin gene. The susceptibility to amphotericin B, itraconazole, voriconazole, posaconazole, ravuconazole, and caspofungin was evaluated according to the Clinical and Laboratory Standards Institute document M38-A2 (CLSI M38-A2) broth microdilution protocol. The results showed that the incident rate of IA was 23.33% and 35 patients with IA (12 proven cases and 23 probable cases) were diagnosed according to the European Organization for Research and Treatment of Cancer and Mycoses Study Group criteria. The 35 patients with IA in the current study comprised 19 men (54.29%) and 16 women (45.71%) with the median age of 42 years. AML (31.5%) was documented as the most prevalent risk factor among our subjects with IA and Aspergillus flavus (65.7%) was the most prevailing causal agent in this study. Among patients with IA, ague (71%) and cough (60%) were the most common symptoms. In the present study, a sensitivity of 94% and a specificity of 98% was reported for GM ELISA in BAL specimens. Also, a sensitivity of 58% and a specificity of 98% was reported for GM ELISA in serum samples. Among 6 tested antifungal drugs, the lowest minimum inhibitory concentration (MIC) values were observed for posaconazole and ravuconazole which showed the range of 0.008-0.0062 μgml and 0.031-0.125 μgml, respectively. The current study has demonstrated that determining

Topics: Adult; Antifungal Agents; Aspergillosis; Bronchoalveolar Lavage Fluid; Cross-Sectional Studies; Female; Galactose; Hematologic Neoplasms; Humans; Immunoenzyme Techniques; Male; Mannans; Organ Transplantation; Pharmaceutical Preparations; Sensitivity and Specificity

A commercial Aspergillus galactomannan antigen (GMA) enzyme linked immunosorbent assay (ELISA) is used to support a diagnosis of systemic aspergillosis in dogs. In human patients, false positive results have been associated with administration of medications derived from molds. We sought to determine the effect of administration of a commercially available oral probiotic nutraceutical that contained Aspergillus-derived ingredients on serum and urine Aspergillus GMA levels in dogs by conducting a prospective, cross-over study. Galactomannan index (GMI) was measured on the solubilized probiotic nutraceutical and was positive (GMI ≥ 0.5) with a mean of 7.91. Serum and urine galactomannan indices were measured in 10 healthy dogs before (day 0) and after 1 week (day 7) of probiotic nutraceutical administration, then again 2 weeks after the probiotic nutraceutical was discontinued (day 21). Median (range) serum GMI were 0.19 (0.08-0.62), 0.22 (0.07-1.15) and 0.17 (0.14-0.63) at day 0, 7 and 21, respectively. Two of 10 dogs developed positive GMI (≥0.5) results after probiotic nutraceutical administration; however, no significant changes were noted over the study period. Median (range) urine GMI results were 0.06 (0.04-0.22), 0.07 (0.05-0.41) and 0.06 (0.03-0.16) at day 0, 7 and 21, respectively. A trend towards an increase urine GMI was noted between day 0 and 7 (P = 0.18), and decrease was noted between day 7 and 21 (P = 0.09). Administration of probiotics containing Aspergillus-derived ingredients to dogs did not reliably result in elevated Aspergillus GMA levels.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus; Dietary Supplements; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Male; Mannans; Probiotics

To investigate the accuracy of immunohistochemistry (IHC) tests for distinguishing between mucormycosis and aspergillosis and compare the clinical characteristics of mucormycosis patients according to galactomannan (GM) results.. We evaluated diagnostic performance of IHC test with tissue sections of patients with culture-proven invasive fungal infection. In addition, we conducted PCR assay with tissue sections of mucormycosis patients with positive GM results to evaluate the possibility of co-infection.. In culture-proven mucormycosis (n = 13) and aspergillosis (n = 20), the sensitivity and specificity of IHC test were both 100% for mucormycosis and 85% and 100%, respectively, for aspergillosis. Among the 53 patients who met the modified criteria for proven mucormycosis and had GM assay results, 24 (45%) were positive. Compared with those with negative GM results (n = 29), mucormycosis patients with positive GM results had significantly higher incidence of gastrointestinal tract infections (6/24 [25%] vs 0/29 [0%], P = .006) and were more likely to be histomorphologically diagnosed as aspergillosis (7/24 [29%] vs 2/29 [7%], P = .06). PCR assay amplified both Aspergillus- and Mucorales-specific DNA in 6 of these 24 cases.. Immunohistochemistry tests seem useful for compensating for the limitations of histomorphologic diagnosis in distinguishing between mucormycosis and aspergillosis. Some proven mucormycosis patients with positive GM results had histopathology consistent with aspergillosis and gastrointestinal mucormycosis. In addition, about one quarter of these patients revealed the evidence of co-infection with aspergillosis by PCR assay.

Topics: Adult; Aged; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; DNA, Fungal; Female; Galactose; Humans; Immunohistochemistry; Invasive Fungal Infections; Male; Mannans; Middle Aged; Mucorales; Mucormycosis; Reagent Kits, Diagnostic; Reproducibility of Results; Retrospective Studies; Sensitivity and Specificity

Galactomannan (GM) detection in biological samples has been shown to predict therapeutic response by azoles and polyenes. In a murine invasive pulmonary aspergillosis model, fosmanogepix or posaconazole treatment resulted in an ∼6- to 7-log reduction in conidial equivalents (CE)/g lung tissue after 96 h versus placebo. Changes in GM levels in BAL fluid and serum mirrored reductions in lung CE, with significant decreases seen after 96 h or 72 h for fosmanogepix or posaconazole, respectively (

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Biomarkers; Galactose; Immunocompromised Host; Invasive Pulmonary Aspergillosis; Lung; Male; Mannans; Mice; Microbial Sensitivity Tests; Triazoles

Aspergillus galactomannan immunoassay is a main diagnostic and monitoring tool in medical mycology. However, the specificity of the method can be skewered by the presence of several other fungi. Trying to diagnose a possible fungal infection of the lower respiratory tract in a haematology patient, it appeared that the fungus Trichoderma longibrachiatum is an additional probable cause of positive galactomannan results. Although, that Trichoderma is a rare but emerging pathogen in immunocompromised patients, the above information could be a caution point in the clinical evaluation of diagnostic results.

Topics: Aspergillosis; Aspergillus; Cell Wall; DNA, Ribosomal Spacer; Galactose; Immunoassay; Invasive Fungal Infections; Lymphoma, Non-Hodgkin; Mannans; Trichoderma

Pathologic diagnosis remains the gold standard for final diagnosis of acute invasive fungal sinusitis (AIFS); however, other less invasive tests could suggest the presence of AIFS in at-risk populations where early diagnosis is crucial. Serum galactomannan Aspergillus antigen has been shown to correlate with a diagnosis of invasive pulmonary aspergillosis; however, it has not adequately been evaluated in regard to AIFS. The objective of this study is to evaluate the statistical relevance of galactomannan in predicting diagnosis of AIFS.. This study was a retrospective review of pathologic records using Co-Path from 2006 to 2017, incorporating 2 separate searches with designated criteria identifying patients who received pathologic evaluation for invasive fungal sinusitis. Electronic medical records were subsequently reviewed. After exclusions isolating at-risk populations and removing duplications, 78 cases were reviewed using the indicated search criteria. Of these, 38 met further criteria of having had both pathologic evaluation and galactomannan analysis. Statistical variables were assessed, as well as all-cause mortality. Peak and closest galactomannan levels were evaluated.. Overall, galactomannan had a sensitivity of 44.8% (95% confidence interval [CI], 26.5% to 64.3%), specificity of 100% (95% CI, 66.4% to 100%), positive predictive value of 100% (95% CI, 74.3% to 100%), and negative predictive value of 36% (95% CI, 18.0% to 57.5%). No significant association was observed in galactomannan status and mortality in this patient population.. Positive serum galactomannan can be an indication of AIFS in patients with a high clinical suspicion. In our study, a positive galactomannan always correlated with a positive pathologic diagnosis. However, given its low sensitivity, one must use caution in relying on galactomannan as a screening tool in diagnosis of AIFS.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Aspergillus; Female; Galactose; Humans; Invasive Fungal Infections; Male; Mannans; Middle Aged; Predictive Value of Tests; Retrospective Studies; Sensitivity and Specificity; Sinusitis; Survival Analysis; Young Adult

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage; Cytokines; Galactose; Hematologic Neoplasms; Humans; Invasive Pulmonary Aspergillosis; Mannans; Polymerase Chain Reaction

Invasive pulmonary aspergillosis is a life-threatening fungal disease principally caused by the ubiquitous mould Aspergillus fumigatus. This clinical entity is a major cause of morbidity and mortality (principally, but not restricted to, immunocompromised individuals). A few recent reports suggest in vitro fungicidal activity of sertraline against Aspergillus spp., but this activity has not yet been investigated in vivo.. To evaluate the antifungal activity of sertraline in two in vivo models of aspergillosis.. The antifungal activity of sertraline as monotherapy at three different doses (3, 10 and 15 mg/kg) was evaluated in Galleria mellonella and in a murine model of invasive pulmonary aspergillosis. Therapeutic efficacy parameters determined were larval survival and health index score for G. mellonella, whereas pulmonary fungal burden, galactomannan and lung histopathology were assessed in the murine model.. Sertraline treatments improved larval survival and health index score, especially at doses of 10 and 15 mg/kg. Moreover, 10 mg/kg sertraline was able to reduce pulmonary fungal burden with an efficacy comparable with that of 3 mg/kg amphotericin B and 10 mg/kg voriconazole.. To the best of our knowledge, this is the first in vivo study that evaluates the antifungal activity of sertraline against A. fumigatus, showing a possible promising option for the adjuvant treatment of pulmonary aspergillosis.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Colony Count, Microbial; Disease Models, Animal; Galactose; Histocytochemistry; Lepidoptera; Lung; Male; Mannans; Mice, Inbred BALB C; Sertraline; Survival Analysis; Treatment Outcome

Topics: Antifungal Agents; Aspergillosis; Echocardiography; Fatal Outcome; Female; Galactose; Heart; Humans; Mannans; Middle Aged; Tomography, X-Ray Computed; Ventricular Outflow Obstruction; Voriconazole

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Candidiasis; Disaccharides; Europe; Female; Galactose; Humans; Intersectoral Collaboration; Invasive Fungal Infections; Male; Mannans; Mass Spectrometry; Middle Aged; Mucormycosis; Sensitivity and Specificity; Young Adult

Data on whether positive non-sterile fungal culture has the same clinical value as a positive galactomannan (GM) result are limited.. Patients with biopsy-proven invasive aspergillosis or mucormycosis (over an 8-year period) in whom the results of GM and fungal culture of sputum and/or sinus aspirates were available were enrolled. Biopsy-proven cases were defined if fungal culture from a sterile biopsy specimen gave a positive result and/or hyphae were demonstrated by immunohistochemical staining for aspergillosis and mucormycosis.. A total of 71 patients comprising 30 biopsy-proven cases of aspergillosis including 13 cases with positive sterile cultures and 41 biopsy-proven cases of mucormycosis including eight cases with positive sterile cultures were enrolled. Of 30 patients with aspergillosis, 15 (50%) revealed Aspergillus spp. growth from non-sterile site and none exhibited the agents of mucormycosis growth from non-sterile site. However, of 41 patients with mucormycosis, eight (20%) revealed the agents of mucormycosis growth from non-sterile site and three (7%) exhibited Aspergillus spp. growth from non-sterile site. In terms of GM assays, 23 (77%) of 30 patients with aspergillosis revealed positive GM results, and 17 (41%) of 41 patients with mucormycosis revealed positive GM assays. So, positive fungal culture from non-sterile site (88% [23/26]) were better correlated with the diagnosis than positive GM assay (57% [23/40]) (p value = .01).. Positive fungal cultures from non-sterile sites better correlate with the diagnosis of aspergillosis and mucormycosis based on sterile culture results and histopathological findings than positive GM results.

Topics: Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Female; Galactose; Humans; Immunohistochemistry; Male; Mannans; Microbiological Techniques; Middle Aged; Mucorales; Mucormycosis; Paraffin Embedding; Republic of Korea; Sputum; Sterilization; Tertiary Care Centers; Young Adult

The incidence of false-positive serum galactomannan (GM) enzyme-linked immunosorbent assay test results has been scarcely examined among older subjects. Additionally, previous studies have highlighted the influence of serum immunoglobulin G (IgG) levels on GM test results. We hypothesised that age-related IgG level elevation might also be associated with false-positive GM test results in older subjects.. This study aimed to examine the association between false-positive GM test results and age or serum IgG levels.. We investigated the association between false-positive serum GM test results and age in 1071 healthy adult subjects. Then, we validated this association and further explored the correlation with serum IgG levels by retrospectively identifying 700 patients with newly diagnosed haematologic disorders without probable or proven invasive aspergillosis.. Healthy subjects with false-positive GM test results were significantly older than those without false-positive results (P < 0.001). Among patients with haematologic disorders, IgG myeloma patients showed significantly higher false-positive rates (57/125 [45.6%]) than patients with other haematologic disorders (non-Hodgkin lymphoma: 48/315 [15.2%], myelodysplastic syndrome/aplastic anaemia: 19/141 [13.5%]; both P < 0.001) or other types of myeloma (IgA myeloma: 13/60 [21.7%], light chain myeloma: 9/52 [17.3%]; both P < 0.01). Furthermore, among non-multiple myeloma patients, advanced age and higher IgG level were also significantly associated with the high frequency of false-positive GM test results.. False-positive serum GM test results were frequent among older subjects and patients with elevated serum IgG levels. These results suggested that age- and/or disease-related IgG level elevation could induce this phenomenon.

Topics: Adult; Age Factors; Aged; Antigens, Fungal; Aspergillosis; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Immunoglobulin G; Male; Mannans; Middle Aged; Retrospective Studies

We report a case of disseminated histoplasmosis in a renal transplant recipient who presented with a nodular pulmonary lesion and elevated serum and bronchoalveolar lavage (BAL) Aspergillus galatomannan. This almost led to an erroneous diagnosis of invasive aspergillosis since the donor respiratory tract was known to be colonized with Aspergillus terreus. However, distinctive intracelluar Histoplasma yeasts on peripheral blood smear led to early diagnosis and appropriate treatment. The cross-reactivity between Aspergillus galactomannan and Histoplasma antigen is discussed further.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Azure Stains; Blood; Bronchoalveolar Lavage Fluid; Female; Galactose; Histoplasma; Histoplasmosis; Humans; Kidney Transplantation; Mannans; Middle Aged; Transplant Recipients

Topics: Animals; Antibodies, Fungal; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus; Aspergillus flavus; Aspergillus fumigatus; Cell Wall; Cross Reactions; Disease Models, Animal; Epitopes; Female; Galactose; Immunologic Tests; Mannans; Mice; Mice, Inbred BALB C; Polysaccharides, Bacterial; Recombinant Proteins

Aspergillus fumigatus frequently colonizes the airways of patients with cystic fibrosis (CF) and may cause various severe infections, such as bronchitis. Serological data, sputum dependent markers and longitudinal data of treated cases of Aspergillus bronchitis were evaluated for further description of this infection. This study, which comprises three substudies, aimed to analyze epidemiological data of Aspergillus in CF and the entity of Aspergillus bronchitis. In a first step, data of the German Cystic Fibrosis Registry were used to evaluate the frequency of Aspergillus colonization in patients with CF (n = 2599). Then a retrospective analysis of 10 cases of Aspergillus bronchitis was performed to evaluate longitudinal data for lung function and clinical presentation parameters: sputum production, cough and physical capacity. Finally, a prospective cohort study (n = 22) was conducted to investigate serological markers for Aspergillus bronchitis: total serum IgE, specific serum IgE, specific serum IgG, as well as sputum galactomannan, real-time PCR detection of Aspergillus DNA in sputum and fungal cultures. Analysis of the German CF registry revealed an Aspergillus colonization rate of 32.5% among the 2599 patients. A retrospective data analysis of 10 treated cases revealed the clinical course of Aspergillus bronchitis, including repeated positive sputum culture findings for A. fumigatus, no antibiotic treatment response, total serum IgE levels <200 kU/l, no observation of new pulmonary infiltrates and appropriate antifungal treatment response. Antifungal treatment durations of 4 ± 1.6 (2-6) weeks significantly reduced cough (P = 0.0067), sputum production (P < 0.0001) and lung function measures (P = 0.0358) but not physical capacity (P = 0.0794). From this retrospective study, a prevalence of 1.6% was calculated. In addition, two cases of Aspergillus bronchitis were identified in the prospective cohort study according to immunological, molecular and microbiological parameters. A prevalence of 9% was assessed. Aspergillus bronchitis appears to occur in a minority of colonized CF patients. Antifungal treatment may reduce respiratory symptoms and restore lung function.

Topics: Adolescent; Adult; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Biomarkers; Bronchitis; Child; Cystic Fibrosis; DNA, Fungal; Female; Galactose; Germany; Humans; Longitudinal Studies; Male; Mannans; Prevalence; Prospective Studies; Real-Time Polymerase Chain Reaction; Respiratory Function Tests; Retrospective Studies; Serum; Sputum; Young Adult

This study aimed to evaluate new tools to diagnose invasive aspergillosis (IA) directly from broncho-alveolar lavage (BAL) samples.. All consecutive patients with suspected IA who underwent bronchoscopy with BAL were prospectively included. Mycological culture and ELISA detection of galactomannan (GM) were performed on BAL. Two in-house and two marketed PCR assays were used on BAL DNA extracts to detect Aspergillus species. Susceptibility testing was performed after culture; marketed PCR assays detected mutations in the CYP51A gene associated to resistance.. Within 3 years, 1555 BAL samples were processed, including 413 samples from 387 immunosuppressed patients. IA diagnosis was no-IA, possible, probable or proven IA in 326, 23, 37 and 1 patients, respectively. PCR assays sensitivity for Aspergillus detection ranged from 61% to 74%, below GM (87%), but contrasting with 47% for cultures. Combining PCR to EORTC/MSG criteria increased the sensitivity to 100%. Interestingly, tests performance in non-hematological patients ranged from 60% to 75%, and were higher than in hematological patients, and those with prior exposure to antifungals. All 16 isolates of Aspergillus fumigatus were susceptible; PCR did not detect any resistance marker in the 37 A. fumigatus PCR-positive samples.. The molecular detection of Aspergillus directly in BAL samples greatly improved the diagnosis of IA, particularly in non-hematological patients.

Topics: Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Cytochrome P-450 Enzyme System; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Fungal Proteins; Galactose; Humans; Immunocompromised Host; Invasive Fungal Infections; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Prospective Studies; Sensitivity and Specificity

Objective This study retrospectively evaluated fungal dissemination due to hospital reconstruction and explored effective methods of predicting an outbreak. Methods Patients suspected of having invasive aspergillosis were tested for Aspergillus galactomannan antigen before and after reconstruction, and the mean values of three months of testing for positive patients were determined. The characteristics of patients with aspergillosis during this period were also assessed. Results Forty-five patients were positive for Aspergillus antigen (>0.5 cut-off index) from January 2013 to December 2014. Mean Aspergillus antigen values significantly increased following reconstruction (p<0.05). Three patients developed pneumonia due to Aspergillus and were diagnosed with "probable" invasive aspergillosis according to the European Organization for Research and Treatment of Cancer and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. We also discovered that the anteroom to contain dust was not prefabricated and a negative pressure system to remove dust was not used. After construction of the unit, no new cases of aspergillosis were diagnosed. Conclusion Many Aspergillus spores may be transiently floating during hospital reconstruction. Therefore, monthly surveillance with frequent serum galactomannan antigen testing to predict outbreaks is necessary. Surveillance of all patients in the hematology ward is especially important. Reconsideration of prophylactic antifungals may also be necessary during hospital reconstruction.

Topics: Adult; Aged; Aged, 80 and over; Air Pollutants; Antigens, Fungal; Aspergillosis; Aspergillus; Disease Outbreaks; Environmental Exposure; Female; Galactose; Hospital Design and Construction; Humans; Male; Mannans; Middle Aged; Retrospective Studies; Sensitivity and Specificity

The European Society for Clinical Microbiology and Infectious Diseases, the European Confederation of Medical Mycology and the European Respiratory Society Joint Clinical Guidelines focus on diagnosis and management of aspergillosis. Of the numerous recommendations, a few are summarized here. Chest computed tomography as well as bronchoscopy with bronchoalveolar lavage (BAL) in patients with suspicion of pulmonary invasive aspergillosis (IA) are strongly recommended. For diagnosis, direct microscopy, preferably using optical brighteners, histopathology and culture are strongly recommended. Serum and BAL galactomannan measures are recommended as markers for the diagnosis of IA. PCR should be considered in conjunction with other diagnostic tests. Pathogen identification to species complex level is strongly recommended for all clinically relevant Aspergillus isolates; antifungal susceptibility testing should be performed in patients with invasive disease in regions with resistance found in contemporary surveillance programmes. Isavuconazole and voriconazole are the preferred agents for first-line treatment of pulmonary IA, whereas liposomal amphotericin B is moderately supported. Combinations of antifungals as primary treatment options are not recommended. Therapeutic drug monitoring is strongly recommended for patients receiving posaconazole suspension or any form of voriconazole for IA treatment, and in refractory disease, where a personalized approach considering reversal of predisposing factors, switching drug class and surgical intervention is also strongly recommended. Primary prophylaxis with posaconazole is strongly recommended in patients with acute myelogenous leukaemia or myelodysplastic syndrome receiving induction chemotherapy. Secondary prophylaxis is strongly recommended in high-risk patients. We strongly recommend treatment duration based on clinical improvement, degree of immunosuppression and response on imaging.

Topics: Antibodies, Fungal; Antifungal Agents; Aspergillosis; Aspergillus; Biopsy; Bronchoalveolar Lavage; Disease Management; Early Diagnosis; Flucytosine; Galactose; Humans; Immunocompromised Host; Immunologic Tests; Invasive Pulmonary Aspergillosis; Itraconazole; Leukemia, Myeloid, Acute; Magnetic Resonance Imaging; Mannans; Microbial Sensitivity Tests; Myelodysplastic Syndromes; Nitriles; Pyridines; Tomography, X-Ray Computed; Triazoles; Voriconazole

Invasive aspergillosis is a major concern in neutropenic patients. We studied the utility of Galactomannan antigen detection test in serum using ELISA technique for early detection of invasive aspergillosis. Diagnostic accuracy of Galactomannan index (GMI) test was maximum at a cut-off of > 1.5 with a negative predictive value of more than 95%.

Topics: Aspergillosis; Chemotherapy-Induced Febrile Neutropenia; Child; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Immunocompromised Host; Mannans; ROC Curve

Topics: Aspergillosis; Aspergillus; Breath Tests; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunocompromised Host; Mannans

This study was conducted to get a complete clinical and mycological picture of invasive aspergillosis (IA) in respiratory medicine ICU of a tertiary care hospital.. From the cohort of 235 patients only one had proven IA. Based on AspICU algorithm, 21 had putative IA (8.9%), 12 were colonised (5.1%).. Adjusting the confounding factors, significant risk factors for IA were chronic obstructive pulmonary disease (COPD), temperature of ≥38°C, pneumonia and acute respiratory distress syndrome (ARDS). The best predictor of IA was AspICU algorithm (AUC, 1) followed by serum galactomannan antigen (GM) cut-off (≥1.24) calculated based on AspICU algorithm (AUC, 0.822). For 37% of patients, IA diagnoses was made earlier with serum GM than radiology. There were 70/235 (29.8%) deaths within 30 days of enrolment in the study. Aspergillus culture positivity (34/235, 14.5%) was associated with very high mortality (27/34, 79.4%), (p<0.05). The best predictor of mortality was GM cut-off (≥1.24) calculated based on AspICU algorithm (AUC, 0.835).. This study imparts the focus on relatively underestimated Aspergillus infections prevalent in ICUs. The AspICU algorithm was found to be useful over others for IA diagnosis. The prognostic usefulness of serum GM antigen detection test highlighted overlooking the same may not be rewarding for the outcome of IA suspected ICU subpopulation.

Topics: Adult; Algorithms; Aspergillosis; Biomarkers; Cross-Sectional Studies; Early Diagnosis; Female; Fever; Galactose; Humans; Intensive Care Units; Male; Mannans; Middle Aged; Pneumonia; Prognosis; Respiratory Distress Syndrome; Risk Factors

To compare the epidemiology, clinical presentation, diagnosis, treatment, and outcome of haematologic patients with invasive aspergillosis (IA) or invasive fusariosis (IF).. We retrospectively reviewed the charts of 36 patients with IA and 26 with IF diagnosed between 2006 and 2017 in haematologic patients, and compared baseline characteristics, coexisting exposures, clinical manifestations, treatment, and the outcome.. Fever was more frequent in IF (96.2% vs. 63.9%, p 0.003), whereas pneumonia (88.9% vs. 50.0%, p 0.001) and sinusitis (63.9% vs. 38.5%, p 0.048) were more frequent in IA. Skin lesions and positive blood cultures occurred exclusively in patients with IF. Among patients with pneumonia, the halo sign was more frequent in IA (62.5% vs. 23.1%, p 0.02). Serum galactomannan was positive in 88.6% of patients with IA and in 73.3% with IF (p 0.18), with no differences in the median number of positive tests and galactomannan values. Positive serum galactomannan plus lung infiltrates was the predominant clinical presentation in IA and occurred in four of 13 patients with IF and lung involvement. The 30-day survival was 77.7% in IA and 46.1% in IF (p 0.01).. IA and IF share the same epidemiologic scenario but different clinical presentations in the majority of cases, with disease in the airways in IA, and fever, metastatic skin lesions, and positive blood cultures in IF. However, a substantial proportion of patients with IF present with a clinical picture similar to IA, with fever, lung infiltrates, and positive serum galactomannan.

Topics: Adult; Aged; Aspergillosis; Female; Fever; Fusariosis; Galactose; Humans; Invasive Fungal Infections; Male; Mannans; Middle Aged; Pneumonia; Retrospective Studies; Young Adult

Invasive aspergillosis (IA) remains a critical issue in immunosuppressed patients. Detection of galactomannan antigen (GM) in serum samples is included as a criterion of IA by the European Organization for the Research and Treatment of Cancer/Mycoses Study Group. Nevertheless, Aspergillus DNA detection by polymerase chain reaction (PCR) has not yet been included because clinical data validation is lacking. The present study describes the simultaneous performance of GM and PCR tests as routine methods for IA diagnosis.. During the period January 2012 to December 2017, a total of 156 white children hospitalized in a tertiary children's hospital of Athens (97 boys and 59 girls; age range, 5 months-14 years) were examined as possible cases of IA. Patients were classified into 4 groups based on their underlying diseases: hematologic malignancies (107 of 156 [68.6%]), solid tumors (16 of 156 [10.2%]), primary immunodeficiency (12 of 156 [7.7%]), and hereditary immunodeficiency (21 of 156 [13.5%]). GM detection was made with the Platelia Aspergillus Ag kit (Bio-Rad Laboratories, Hercules, California). Sera with a cut-off index ≥0.5 on at least 2 separate blood collections were considered positive. Serum detection of Aspergillus DNA was conducted with real-time PCR MycAssay Aspergillus assay (Myconostica Ltd, Cambridge, United Kingdom). PCR positivity was determined by using a threshold of 38 cycles in at least 1 serum sample. Four or more successive samples per patient were tested.. Overall, 28 of 156 patients (53 of 744 serum samples) were found positive. Eleven patients were positive using both methods (24 samples). Four children were positive only by PCR (6 samples), whereas 13 (23 samples) were positive only with GM in consecutive samples. Agreement of both methods, GM(+)/PCR(+) or GM(-)/PCR(-), was found in 139 patients (90% of total patients) and 715 samples (96.1% of total samples). The agreement of both methods was found: (1) 85% in patients with hematologic malignancies; (2) 100% in patients with solid tumors; (3) 97.5% in patients with primary immunodeficiency; and (4) 98.8% in patients with hereditary immunodeficiency. Overall disagreement was observed in 17 patients, in which the positive result in any of the 2 methods was estimated as true positive in conjunction with radiologic and other clinical findings.. The combination of GM and PCR, provided high diagnostic accuracy in consecutive samples (twice a week). Clinical, radiologic, and other laboratory findings should be taken into consideration in the evaluation of GM and PCR.

Topics: Adolescent; Antigens; Aspergillosis; Aspergillus; Child; Child, Preschool; DNA, Fungal; Female; Galactose; Greece; Humans; Immunocompromised Host; Immunologic Deficiency Syndromes; Infant; Male; Mannans; Neoplasms; Real-Time Polymerase Chain Reaction

Galactomannan antigen (GM) testing has been used for decades to screen immunocompromised patients for invasive aspergillosis (IA). Recent publications suggested that using a higher cut-off value than 0.5 in bronchoalveolar lavage fluid (BALF) could be more discriminant for hematology patients. We retrospectively analyzed the values of GM in BALF over 7 years (from 2010 to 2016). Performance indicators of the GM in BALF, according to three different cut-off values (0.5, 0.8, 1.5), were calculated using Stata 14.1. IA classification for hematology patients was based on European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria, as defined in 2008. A number of 716 GM were performed on BALF from 2010 to 2016 (597 patients) and 66 were positive (> 0.5). Among these 597 patients, 27 IA were diagnosed, 13 with a positive GM in BALF, 9 with a negative GM in BALF, and 5 unclassified IA (ICU patients). The analysis of performance indicators, based on our local data, did not demonstrate any significant difference using a higher cut-off value of GM in BALF. This result may be explained by the local recruitment of patients and by pre-analytic variations during BALF realization.

Topics: Aspergillosis; Aspergillus; Biomarkers; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunocompromised Host; Mannans; Reproducibility of Results; Retrospective Studies; Sensitivity and Specificity

Aspergillus terreus causes invasive aspergillosis (IA) in immunocompromised patients. Treatment is complicated by intrinsic resistance to amphotericin B and thereby contributing to a high mortality. Therefore, we conducted in vitro studies to investigate the effectivity of adjunctive recombinant interferon-γ immunotherapy. We describe a pediatric patient with A. terreus IA who received adjunctive recombinant interferon-γ (rIFNγ) immunotherapy. In vitro studies were conducted to investigate the capacity of rIFNγ to improve antifungal host defense in terms of fungal killing ability and the release of pro-inflammatory cytokines in cells of the patient as well as healthy controls. An 8-year-old female pediatric patient with leukemia developed A. terreus IA. She clinically deteriorated and had high serum galactomannan levels despite broad antifungal therapy. Therefore, adjunctive immune stimulatory therapy with rIFNγ was initiated. After 3 weeks of treatment, galactomannan levels decreased and the patient clinically showed improvement. Addition of rIFNγ boosted the capacity of monocytes of healthy volunteers to mount TNFα and IL-1β cytokine responses to Escherichia coli LPS, and increased TNFα response to both A. terreus and Aspergillus fumigatus. Monocytes isolated from the patient's blood demonstrated a similar augmented cytokine induction in response to rIFNγ. In addition, rIFNγ increased the capacity of monocytes from healthy volunteers as well as monocytes from the patient to kill A. terreus spores. Adjuvant immunotherapy with rIFNγ might be a promising additional treatment strategy that could be used to improve outcome in patients with refractory invasive A. terreus infections or other resistant invasive Aspergillus infections.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Cells, Cultured; Child; Cytokines; Female; Galactose; Humans; Immunotherapy; Interferon-gamma; Mannans; Monocytes; Opportunistic Infections; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Recombinant Proteins; Treatment Outcome

Interactions between fungal pathogens such as Aspergillus fumigatus with host alveolar epithelium and innate immune cells are crucial in the defense against opportunistic fungal infections. In this study a simplified Transwell

Topics: A549 Cells; Alveolar Epithelial Cells; Aspergillosis; Aspergillus fumigatus; Cell Membrane Permeability; Coculture Techniques; Cytokines; Dendritic Cells; Galactose; Host-Pathogen Interactions; Humans; Immunity, Innate; L-Lactate Dehydrogenase; Mannans; Models, Immunological; Primary Cell Culture

Topics: Aspergillosis; Child; Fever; Galactose; Humans; Mannans

Topics: Aspergillosis; Child; Fever; Galactose; Humans; Mannans

Herein we describe a 43 year-old Caucasian female patient with acute myeloid leukemia that developed an unconventional form of invasive Aspergillosis. For therapeutic reasons, a Groshong-type central venous catheter was positioned. Monitoring the patient's clinical conditions, positive values for C-reactive protein and galactomannan were correlated with a probably Aspergillosis. Surprisingly no pulmonary evidences were observed. Due to worsening conditions, she was re-hospitalized and a blood culture was performed, whom positivity resulted as the first clinical evidence of Aspergillus fumigatus. Further evidence about species identification was obtained by sequencing the fungal ITS region. We support the clinical value of blood culture as a decisive factor to improve the diagnosis of catheter-related Aspergillosis.

Topics: Adult; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Blood Culture; C-Reactive Protein; Catheter-Related Infections; Culture Media; DNA, Fungal; DNA, Ribosomal Spacer; Female; Galactose; Humans; Leukemia, Myeloid, Acute; Mannans; Neutropenia; Sequence Analysis, DNA; Treatment Outcome

Topics: Adult; Antifungal Agents; Aspergillosis; Aspergillus; Brain; Cerebral Hemorrhage; Female; Galactose; Humans; Lupus Erythematosus, Systemic; Lymph Nodes; Mannans; Thyroid Gland; Tomography, X-Ray Computed; Voriconazole

We found significant correlation between the incidence of severe influenza and Aspergillus antigenemia among medical intensive care unit patients for 7-month observation (coefficient γ=0.976, p<0.001). High-level ambient pollution was noticed for 2 months before the epidemic, highlighting that influenza patients might coinfect with aspergillosis in the community.

Topics: Adult; Aged; Aged, 80 and over; Air Pollution; Antigens, Fungal; Aspergillosis; Aspergillus; Coinfection; Female; Galactose; Humans; Incidence; Influenza, Human; Male; Mannans; Middle Aged; Particulate Matter; Risk Factors; Taiwan

Topics: Acetylcysteine; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Dithiothreitol; Enzyme-Linked Immunosorbent Assay; Expectorants; Galactose; Humans; Mannans; Respiratory Tract Infections; Retrospective Studies

The aim of this study was to determine the prevalence of invasive aspergillosis (IA) in patients with liver cirrhosis and the performance of serum galactomannan (GM) screening. Patients with decompensated liver cirrhosis and patients with compensated liver cirrhosis presenting with fever and/or respiratory symptoms were prospectively enrolled. All patients were screened by serum GM twice weekly irrespective of clinical signs and symptoms. Positive serum GM triggered work-up consisting of chest computed tomography and in case of pathological findings bronchoscopy. 150 patients were included in the study. Two (1.3%) had probable, one (0.7%) had possible, and 147 (98%) had no evidence of IA. Both patients with probable IA had compensated liver cirrhosis. Sensitivity for serum GM screening for probable versus no IA was 0.5 (95% CI, 0.09-0.91), specificity 0.97 (95% CI: 0.92-0.99), negative predictive value 0.99 (95% CI, 0.96-0.99) and positive predictive value (PPV) 0.17 (95% CI, 0.01-0.64). PPV was 0.5 (95% CI, 0.03-0.98) in patients with clinical suspicion of IA. In conclusion, prevalence of IA in patients with liver cirrhosis seems to be low. Targeted GM testing in case of clinical suspicion of IA may be associated with markedly higher PPVs when compared to universal GM screening in patients with liver cirrhosis.

Topics: Aged; Aspergillosis; Cohort Studies; Female; Galactose; Humans; Kaplan-Meier Estimate; Liver Cirrhosis; Male; Mannans; Middle Aged; Prevalence; Prospective Studies; Sensitivity and Specificity

To investigate the performance of the routine serum galactomannan (sGM) assay in the diagnosis of invasive aspergillosis (IA) in high-risk haematology patients receiving prophylaxis with micafungin.. Retrospective study including all haematological patients who received prophylaxis with micafungin during high-risk IA episodes (neutropenic patients after chemotherapy for acute myeloid leukaemia/myelodysplastic syndrome; allogeneic haematopoietic stem-cell transplantation during early neutropenic phase or graft-versus-host disease requiring high prednisone doses) and for whom at least one sGM result was available. Episodes were classified as follows: true-positive (positive GM in the context of IA), false-positive (positive GM result in patients who had no evidence of IA), true-negative (negative GM test results and no IA), or false-negative (negative GM test in the context of IA). Non-evaluable patients were excluded.. Among 146 evaluable episodes, four were true-positive in the context of probable breakthrough IA (incidence of breakthrough IA, 2.7%); 111/146 high-risk episodes (76%) were considered true-negative and 31/146 (21.2%) were considered false-positive. No false-negative episodes were detected. All but one of the false-positive episodes were detected in surveillance GM tests, leading to high-resolution CT scans in eight cases (8/31; 25.8%), all of which were negative. The positive predictive and negative predictive values of sGM for surveillance and diagnostic approaches were 3.2% (1/31) and 100% (110/110) and 75% (3/4) and 100% (1/1), respectively.. Surveillance of asymptomatic patients receiving prophylaxis with micafungin using sGM is unnecessary, because the results are either negative or false-positive. However, sGM remains useful in the diagnosis of breakthrough IA in symptomatic patients during prophylaxis.

Topics: Adult; Antibiotic Prophylaxis; Aspergillosis; Echinocandins; Female; Galactose; Hematologic Neoplasms; Humans; Lipopeptides; Male; Mannans; Micafungin; Retrospective Studies

Aspergillus is a medically important fungal genus that causes a life-threatening infection known as aspergillosis in immunocompromised patients. β-1,3-Glucan is detected in the plasma of patients with aspergillosis and appears to be useful for the diagnosis of aspergillosis. In this study, we cultured Aspergillus spp. in a chemically defined liquid medium and prepared an Aspergillus water-soluble fraction (ASWS) from the culture supernatants. ASWS was found to be primarily composed of polysaccharides and proteins. Nuclear magnetic resonance analysis suggested that ASWS is a complex carbohydrate, consisting of α-1,3-glucan, β-1,3-glucan, galactomannan, and protein. The ASWS from Aspergillus fumigatus showed limulus factor G activity, whereas zymolyase-treated ASWS did not. ASWS was eliminated from the blood more rapidly than Aspergillus solubilized cell wall β-glucan. We analyzed the reactivity of human immunoglobulin towards ASWS by an enzyme-linked immunosorbent assay. Anti-ASWS antibodies were detected in human sera, with titers differing among individuals. This study demonstrated that the ASWS corresponds to the limulus factor G-activating substance found in the blood of patients with aspergillosis.

Topics: Animals; Antibodies, Fungal; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Biomarkers; Fungal Proteins; Galactose; Glucans; Humans; Immunoglobulins; Male; Mannans; Mice, Inbred DBA; Solubility; Water

Topics: Adult; Antifungal Agents; Aspergillosis; Aspergillus; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Bronchoscopy; Female; Galactose; Hematologic Neoplasms; Humans; Invasive Pulmonary Aspergillosis; Male; Mannans; Middle Aged; Tomography, X-Ray Computed

In recent years galactomannan antigen testing (GM) and also Aspergillus PCR have become increasingly important for diagnosis of invasive aspergillosis (IA). Whether or not these tests need to be performed with bronchoalveolar lavage fluid (BALF; i.e., primary site of infection), or testing of blood samples is sufficient, remains, however, a matter of debate. We evaluated the diagnostic performance of GM ELISA, and Aspergillus PCR by using BALF samples and blood samples obtained at the same day from a total of 53 immunocompromised patients (16 with probable/proven IA and 37 with no evidence of IA according to the revised EORTC/MSG criteria; 38 patients with hematological malignancies were prospectively enrolled at the Medical University of Graz, Austria, 15 patients with mixed underlying diseases at the Mannheim University Hospital). Patients with possible IA were excluded from this analysis. A total of 34/53 (64%) of all patients and 12/16 (75%) of patients with probable/proven IA received mold-active antifungal prophylaxis/therapy at the time of the BALF procedure. Sensitivities of GM and Aspergillus PCR were 38% and 44% in BALF, and 31% and 0% in blood, respectively. Best sensitivity (75%) for detecting proven/probable IA was achieved when BALF Aspergillus PCR, BALF GM (>1.0 ODI), BALF-culture and serum-GM (>0.5 ODI) were combined (specificity 95%). In conclusion, sensitivities of the evaluated diagnostic tests-when interpreted on their own-were low in BALF and even lower in blood, sensitivities increased markedly when diagnostic tests were combined.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; DNA, Fungal; Female; Galactose; Humans; Immunocompromised Host; Invasive Fungal Infections; Male; Mannans; Microbiological Techniques; Middle Aged; Molecular Diagnostic Techniques; Polymerase Chain Reaction; Sensitivity and Specificity

Invasive aspergillosis (IA) is associated with a high morbidity and mortality. Since Aspergillus species are usually not cultured in these patients, presumptive diagnosis of IA is more commonly based on galactomannan (GM) detection. Several factors are known to cause false-positive results in the GM test, but little is known on the influence of pre-analytical variables interfering on the test. Here we studied the influence of temperature and sample storage duration in GM results, using samples known to be negative and positive (spiked) for GM. We also evaluated the effect of hemolysis and hyperbilirubinemia on GM optical indexes. We found no influence of storage time (up to 96 h) and temperatures (refrigerated vs. RT) on GM results. However, bilirubin (P = .022) and haemoglobin (P = .003) content influenced GM readings in samples known for being GM positive and negative at baseline, respectively. We conclude that the Platelia GM test does not suffer major influence of pre-analytical variables such as storage conditions, and low levels of hemolysis and hyperbilirubinemia. Nonetheless, massive haemolysis seems to interfere with GM readings in GM-negative samples, and high levels of bilirubin can affect GM readings in samples that are positive for GM at baseline. These findings may facilitate logistics and the implementation of standard operational procedures in clinical laboratories.

Topics: Adult; Aspergillosis; Bilirubin; Blood Chemical Analysis; False Positive Reactions; Female; Galactose; Hemoglobins; Humans; Male; Mannans; Specimen Handling; Temperature; Time Factors; Young Adult

Pseudallescheria boydii is a fungal organism known to affect immunocompromised patients. This organism is known to cause, in severe cases, invasive infection of various organs such as the central nervous, cardiovascular, and respiratory systems. We report an unusual case of pulmonary P. boydii pneumonia in an immunocompromised critically ill patient with a co-infection of Aspergillus fumigatus and Aspergillus terreus with ARDS. This case highlights the importance of a high index of suspicion for superimposed fungal infections in patients who are critically ill and immunocompromised. Uncommon fungal pathogens should be considered in the differential diagnosis of respiratory failure, especially if diagnostic markers such as galactomannan (from BAL and serum) or 1,3-beta-D-glucan are elevated. Further diagnostic interventions are warranted when insufficient clinical improvement is observed to prevent treatment failure and adverse outcomes.

Topics: Aged; Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Clarithromycin; Coinfection; Critical Illness; Extracorporeal Membrane Oxygenation; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Linezolid; Male; Mannans; Meropenem; Pneumonia; Pseudallescheria; Severe Acute Respiratory Syndrome; Thienamycins; Transplant Recipients; Voriconazole

Rapid diagnosis and early treatment of invasive aspergillosis is crucial for the management of the patients with haematological malignancy. We evaluated 358 sera from 78 febrile neutropenic episodes in patient with invasive aspergillosis (IA) (one proven, 17 probable, and 60 possible) and 83 episodes in patients with no IA according to the EORTC/MSG criteria. Patient's specimens were tested by Mycassay Aspergillus PCR (first commercial real-time PCR test) and in house real-time PCR to investigate the presence of Aspergillus DNA, and by ELISA for detect the galactomannan (GM) antigen. We systematically investigated the medical background that can be effective on the test results. The hospitalisation period was longer in proven/probable episodes when compared with no IA (P = 0.001) and possible episodes. With regard to duration of neutropenia, the differences between both proven/probable with no IA (P = 0.023) and possible with no IA (P = 0.002) were highly significant. Similarly, the rates of T cell suppressant therapy in group proven/probable and possible episodes were significantly higher than in no IA (P = 0.005). There are significant differences in the performance of GM and PCR-based tests among studies, and standardisation is required. Therefore, it can be useful to determine the effective factors on these tests. The use of larger volume of sera improved the performance of real-time PCR for detection of Aspergillus DNA in high-risk adult patients in the present study. Some host factors such as duration of neutropenia and administration of T cell suppressants related to the development of IA.

Topics: Adult; Aspergillosis; Aspergillus; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Fever; Galactose; Hematologic Neoplasms; Humans; Immunosuppressive Agents; Male; Mannans; Middle Aged; Neutropenia; Real-Time Polymerase Chain Reaction

Galactomannan (GM) testing of urine specimens may provide important advantages, compared to serum testing, such as easy noninvasive sample collection. We evaluated a total of 632 serial urine samples from 71 patients with underlying hematological malignancies and found that the urine GM/creatinine ratio, i.e., (urine GM level × 100)/urine creatinine level, which takes urine dilution into account, reliably detected invasive aspergillosis and may be a promising diagnostic tool for patients with hematological malignancies. (This study has been registered at ClinicalTrials.gov under registration no. NCT01576653.).

Topics: Aspergillosis; Biomarkers; Creatinine; Female; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Reproducibility of Results; ROC Curve

Infections are an important cause of morbidity and mortality in patients with rheumatoid arthritis. Patients receiving immunosuppressive or anti-tumor necrosis factor (TNF) agents are vulnerable to fungal infections, including those derived from Aspergillus species. Detection of the Aspergillus galactomannan antigen in serum is useful for the early diagnosis of invasive aspergillosis in patients with hematological malignancies. However, its usefulness for detecting early invasive aspergillosis in rheumatoid arthritis patients remains unestablished.. Galactomannan antigen levels were measured in 340 patients (311 female patients). For patients who exhibited galactomannan antigen levels ≥0.5 during the initial examination, a second examination was performed 3-6 months later. Conventional blood tests and chest radiography were also performed.. Elevated galactomannan antigen levels (≥0.5) were observed in 62 (18.2%) of 340 patients during the initial examination. A second examination was performed in 56 of 62 patients, 50 of whom exhibited elevated antigen levels. Elevated antigen levels were not associated with the use of any drug including anti-TNF agents. Serum galactomannan antigen levels were correlated with the albumin/globulin ratio (r=-0.19, p<0.001), γ-globulin (%; r=0.17, p=0.001), and hemoglobin concentration (r=-0.15, p=0.005). No patient was clinically diagnosed with invasive aspergillosis during the study period.. Serum galactomannan antigen levels are frequently elevated in a nonspecific manner in patients with rheumatoid arthritis.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Arthritis, Rheumatoid; Aspergillosis; Aspergillus; Biomarkers; Female; Galactose; Glucocorticoids; Humans; Hypergammaglobulinemia; Immunosuppressive Agents; Male; Mannans; Middle Aged

False-positive galactomannan (GM) results have been reported in patients treated with gluconate-containing solutions, such as Plasmalyte. The GM optical density index was tested on 33 distinct batches of Plasmalyte and was found to be negative in all of the batches, confirming that Plasmalyte is no longer a cause of false-positive GM results.

Topics: Antigens, Fungal; Aspergillosis; Biomarkers; Electrolytes; False Positive Reactions; Galactose; Humans; Immunoenzyme Techniques; Mannans

Voriconazole is the agent of choice for the treatment of invasive aspergillosis in children at least 2 years of age. The galactomannan index is a routinely used diagnostic marker for invasive aspergillosis and can be useful for following the clinical response to antifungal treatment. The aim of this study was to develop a pharmacokinetic-pharmacodynamic (PK-PD) mathematical model that links the pharmacokinetics of voriconazole with the galactomannan readout in children. Twelve children receiving voriconazole for treatment of proven, probable, and possible invasive fungal infections were studied. A previously published population PK model was used as the Bayesian prior. The PK-PD model was used to estimate the average area under the concentration-time curve (AUC) in each patient and the resultant galactomannan-time profile. The relationship between the ratio of the AUC to the concentration of voriconazole that induced half maximal killing (AUC/EC50) and the terminal galactomannan level was determined. The voriconazole concentration-time and galactomannan-time profiles were both highly variable. Despite this variability, the fit of the PK-PD model was good, enabling both the pharmacokinetics and pharmacodynamics to be described in individual children. (AUC/EC50)/15.4 predicted terminal galactomannan (P= 0.003), and a ratio of >6 suggested a lower terminal galactomannan level (P= 0.07). The construction of linked PK-PD models is the first step in developing control software that enables not only individualized voriconazole dosages but also individualized concentration targets to achieve suppression of galactomannan levels in a timely and optimally precise manner. Controlling galactomannan levels is a first critical step to maximizing clinical response and survival.

Topics: Antifungal Agents; Area Under Curve; Aspergillosis; Aspergillus fumigatus; Biomarkers; Child; Child, Preschool; Computer Simulation; Drug Monitoring; Female; Fungal Polysaccharides; Galactose; Humans; Male; Mannans; Microbial Sensitivity Tests; Models, Statistical; Precision Medicine; Voriconazole

To investigate the diagnosis and treatment of invasive pulmonary aspergillosis (IPA) in children.. The clinical data of 16 cases of proven or probable IPA who had been in our Hospital from January 2006 to June 2014 were retrospectively analyzed.. Among the 16 patients, 11 were males and 5 were females. One child had proven IPA and 15 children had probable IPA. Host risk included long duration use of multiple broad-spectrum antibiotics in 16 cases, neutropenia in 9 cases, invasive mechanical ventilation in 3 cases, primary immunodeficiency disease in 2 cases, long-term use of glucocorticoids in 2 cases, measles in 2 cases, and congenital pulmonary hypoplasia in 1 case. Fever, cough and expectoration were present in all the children with IPA. At the time of diagnosis, the halo sign and subpleural wedge consolidation shadows were more common in neutropenia group (5/9, 7/9) than those in non-neutropenia group(0/7, 1/7)(P<0.05). The cavities and"air-crescent sign"were more common after 15 days to 1 month when the children had been treated with anti-aspergillosis drugs than that at the onset of diagnosis of IPA (P<0.05). The positive rate of serum galactomannan (GM) test was higher than that of sputum culture and serum G test (P<0.05). Thirteen children received voriconazole, in 7 of the children the treatment was effective.. Neutropenia were the common host risk factors in children with IPA. Subpleural wedge consolidation shadows, the halo sign and the"air-crescent"sign were highly suggestive of the diagnosis of IPA in children. Subpleural wedge consolidation shadows and the halo sign were more common in neutropenia group than in non-neutropenia group in the early stage of the course. Serum GM test played an important role in the diagnosis of IPA in children. Voriconazole was effective in majority of the children with IPA.

Topics: Anti-Bacterial Agents; Aspergillosis; Child; Cough; Female; Fever; Galactose; Glucocorticoids; Humans; Male; Mannans; Measles; Neutropenia; Retrospective Studies; Risk Factors; Sputum; Voriconazole

Invasive aspergillosis (IA) has recently increased and has a high mortality rate in immunocompromised patients. IA before hematopoietic stem cell transplantation (HSCT) is not uncommon, but how to cope with it is very tough. The serum aspergillus galactomannan antigen (GM) is a helpful marker for diagnosis of IA, and a serial follow-up of GM levels is important to evaluate the response of treatment. However, data on the changes of GM during HSCT are very limited.. Patient 1 was a 2-year-old female with severe aplastic anemia. A typical lung lesion in the computed tomography of the chest with elevated GM levels was noted, and probable IA was diagnosed. After a combination treatment of voriconazole and caspofungin, the GM levels decreased. Although of significant improvement, the pulmonary lesion in the chest X-ray did not disappear before HSCT. The GM levels increased when she received the conditioning regimen during HSCT. The GM levels remained high during the use of steroids for the graft-versus-host disease and declined gradually after tapering off steroids and cyclosporine. Patient 2 was a 12-year-old female with severe aplastic anemia. Voriconazole was administered after the diagnosis of a probable IA. The pulmonary lesions in the chest X-ray disappeared before HSCT. The GM levels flared up during the administration of conditioning regimen and declined after neutrophil engraftment. At present, the two patients were cured of the disease without requiring surgical resection of their pulmonary IA.. To our knowledge, this is the first report about the changes of GM during HSCT in patients with prior IA. With appropriate antifungal therapy and restoration of patient's immunity, IA can be cured without surgical resection. Further studies are warranted.

Topics: Antifungal Agents; Aspergillosis; Child; Diagnosis, Differential; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Infant; Mannans; Tomography, X-Ray Computed

Here, we report a patient developing a postoperative peritoneal infection by Aspergillus fumigatus. While galactomannan serum levels were negative throughout the time course, galactomannan levels in peritoneal fluids yielded high results. Serological testing of peritoneal fluids for fungal antigens might be a useful and easily applicable tool to support diagnosis of intraabdominal aspergillosis, which represents a rare type of invasive fungal infection.

Topics: Aged; Antigens, Fungal; Ascitic Fluid; Aspergillosis; Aspergillus fumigatus; Candida glabrata; Candidiasis; Galactose; Germany; Humans; Male; Mannans; Peritoneal Cavity; Peritonitis; Postoperative Complications; Thoracic Surgical Procedures

Acute invasive fungal rhinosinusitis (AIFR) is an aggressive opportunistic infection with a high mortality rate. Recently, non-invasive techniques have been introduced for diagnosis of invasive fungal disease. The purpose of this study is to evaluate the diagnostic significance of serum galactomannan measurement in patients with AIFR.. We conducted a retrospective case-control study of 28 patients with AIFR and 36 fungus ball (FB) patients. We evaluated clinical, laboratory, and pathologic findings along with disease course.. In 28 patients with AIFR, there were 21 cases of invasive aspergillosis (IA) and 7 cases of invasive mucormycosis (IM). The control group was comprised of 36 patients with FB. The three-group analysis showed a statistically significant difference among the groups. At the cut-off value of 0.48, the sensitivity and specificity were 71.4% and 93.0%, respectively. Comparison of mean serum galactomannan levels in 5 non-survivors and 9 survivors at initial measurement showed no significant difference, but that became significantly different 1 week later. Statistical analysis showed that the levels of serum galactomannan decreased significantly according to the measurement-point in within survivor-group analysis. The difference in between survivor-groups analysis was also significant.. Serum galactomannan measurement seems useful for early diagnosis and discrimination of fungal species in patients with AIFR. In addition, clinical outcomes may be related to the levels and patterns of serum galactomannan, especially in IA. The appropriate measurement of galactomannan might be helpful in treating the patients at high risk for AIFR.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Case-Control Studies; Child; Female; Galactose; Humans; Invasive Fungal Infections; Male; Mannans; Middle Aged; Mucormycosis; Retrospective Studies; Rhinitis; Sensitivity and Specificity; Sinusitis; Young Adult

High mortality rates of invasive fungal disease (IFD), especially invasive aspergillosis (IA), in immunocompromised haematological patients and current diagnostic limitations require improvement of detection of fungal pathogens by defining the optimal use of biomarkers and clinical samples. Concurrent bronchoalveolar lavage (BAL) and peripheral blood samples of 99 haematological patients with suspected IFD were investigated within a multicentre prospective study. Diagnostic performance of a galactomannan (GM) enzyme immune assay (EIA), a 1,3-β-D-glucan assay (BDG), an Aspergillus PCR, and a multifungal DNA-microarray (Chip) alone or in combination were calculated. IFD were classified as proven (n=3), probable (n=34), possible (n=33), and no IFD (n=29) according to EORTC/MSG criteria. GM, PCR, and Chip showed superior diagnostic performance in BAL than in blood, whereas specificity of BDG in BAL was poor (48% (14/29)). The combination of GM (BAL) with BDG (blood) showed sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and DOR (diagnostic odds ratio) of 92% (34/37), 93% (27/29), 94%, 90%, and 153.0, respectively. Combining GM (BAL) with PCR (BAL) showed convincing diagnostic potential for diagnosing IA with sensitivity, specificity, PPV, NPV, and DOR of 85% (17/20), 97% (28/29), 94%, 90%, and 158.7. Addition of the DNA-microarray resulted in further detection of two mucormycetes infections. In 1 out of 15 Aspergillus DNA-positive samples a triazole resistance-mediating Cyp51A mutation was found. Combination of biomarkers is superior to their sole use in diagnosing IFD, particularly IA. Integrating blood and BAL samples into a diagnostic algorithm is an advantageous approach.

Topics: Aspergillosis; Aspergillus; Azoles; beta-Glucans; Bronchoalveolar Lavage Fluid; Galactose; Humans; Invasive Fungal Infections; Mannans; Microbiological Techniques; Molecular Diagnostic Techniques; Multiplex Polymerase Chain Reaction; Oligonucleotide Array Sequence Analysis; Prospective Studies; Sensitivity and Specificity

Invasive aspergillosis (IA) is a fatal infection that is difficult to diagnose in immunocompromised patients. In this study, Aspergillus-specific DNA was searched using real-time PCR (RT-PCR) in serum samples. Galactomannan (GM) and/or beta-D-glucan (BDG) tests were previously performed on these samples for 70 neutropenic patients with hematological malignancy.. The patients were categorized according to the criteria of the European Organization for Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG). Among the patient serum samples, the first positive GM or BDG test sample and the median sample of GM or BDG test for negative patients were used to detect DNA levels by RT-PCR method (Light Cycler 480, Roche Molecular Biochemicals, Meylan, France) using a commercial kit (Way2Gene Fungi; Genmar, İzmir, Turkey).. When the proven and probable IA group were considered as real patients, sensitivity of Aspergillus-specific DNA test was 90%, specificity was 73.3%, positive predictive value was 81.8%, and negative predictive value was 84.6%.. This study found that searching for specific DNA by RT-PCR method has a sensitivity as high as the GM test. Although specificity was rather low, it was concluded that it can be used jointly with GM and BDG tests after decreasing contamination by severe laboratory applications.

Topics: Adult; Aspergillosis; Aspergillus; DNA; Galactose; Hematologic Neoplasms; Humans; Mannans; Turkey

The galactomannan is a major cell wall molecule of Aspergillus fumigatus. This molecule is composed of a linear mannan with a repeating unit composed of four α1,6 and α1,2 linked mannose with side chains of galactofuran. To obtain a better understanding of the mannan biosynthesis in A. fumigatus, it was decided to undertake the successive deletion of the 11 genes which are putative orthologs of the mannosyltransferases responsible for establishing α1,6 and α1,2 mannose linkages in yeast. These deletions did not lead to a reduction of the mannan content of the cell wall of the mycelium of A. fumigatus. In contrast, the mannan content of the conidial cell wall was reduced and this reduction was associated with a partial disorganization of the cell wall leading to defects in conidial survival both in vitro and in vivo.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Carbohydrate Conformation; Cell Wall; Fungal Proteins; Galactose; Gene Deletion; Gene Expression Regulation, Fungal; Host-Pathogen Interactions; Mannans; Mannose; Mannosyltransferases; Mice; Mycelium; Spores, Fungal; Virulence

We investigated the clinical performance of a polymerase chain reaction (PCR)-based commercial platform, the Myconostica MycAssay™ Aspergillus (MAP), for fungal DNA detection in the serum of patients at risk of invasive aspergillosis (IA). Sixty-four hospitalized patients were prospectively enrolled and a total of 71 different episodes were investigated (30 episodes were clinically/microbiologically classified as IA and 41 as control episodes). When MAP was compared to the galactomannan (GM) assay, no significant differences were found in terms of sensitivity (46.7% vs. 50.0%), specificity (97.6% vs. 95.1%), positive predictive value (PPV) (93.3% vs. 88.2%), and negative predictive value (NPV) (71.4% vs. 72.2%). The corresponding areas under the curve (AUC) of the receiver operating characteristic (ROC) curves were also superimposable. Overall, because of the good agreement between the two assays and considering the high specificity and PPV of the MAP, we suggest the use of this PCR-based platform as a second-level examination for the evaluation of clinically undefined cases where culture or GM have provided positive results.

Topics: Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; DNA, Fungal; Female; Fungemia; Galactose; Humans; Immunoenzyme Techniques; Male; Mannans; Middle Aged; Molecular Diagnostic Techniques; Predictive Value of Tests; Prospective Studies; Real-Time Polymerase Chain Reaction; ROC Curve; Sensitivity and Specificity; Young Adult

Echinocandins inhibit β-1,3-glucan synthesis and are one of the few antimycotic drug classes effective against Aspergillus spp. In this study, we characterized the β-1,3-glucan synthase Fks1 of Aspergillus fumigatus, the putative target of echinocandins. Data obtained with a conditional mutant suggest that fks1 is not essential. In agreement, we successfully constructed a viable Δfks1 deletion mutant. Lack of Fks1 results in characteristic growth phenotypes similar to wild type treated with echinocandins and an increased susceptibility to calcofluor white and sodium dodecyl sulfate. In agreement with Fks1 being the only β-1,3-glucan synthase in A. fumigatus, the cell wall is devoid of β-1,3-glucan. This is accompanied by a compensatory increase of chitin and galactosaminogalactan and a significant decrease in cell wall galactomannan due to a massively enhanced galactomannan shedding. Our data furthermore suggest that inhibition of hyphal septation can overcome the limitations of echinocandin therapy. Compounds inhibiting septum formation boosted the antifungal activity of caspofungin. Thus, development of clinically applicable inhibitors of septum formation is a promising strategy to improve existing antifungal therapy.

Topics: Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Benzenesulfonates; beta-Glucans; Caspofungin; Cell Wall; Chitin; Echinocandins; Galactose; Glucosyltransferases; Hyphae; Lipopeptides; Mannans; Mutation; Phenotype; Polysaccharides

We evaluated the incidence of and risk factors for false-positive Aspergillus galactomannan (GM) antigenemia in allogeneic hematopoietic stem cell transplantation (HSCT). We also focused on the GM index value and its kinetics.. Patients who underwent their first allogeneic HSCT at our center between June 2007 and December 2012 were included (n = 172). Episodes of positive GM tests were classified as either "true-positive", which fulfilled the EORTC criteria for proven or probable invasive aspergillosis (IA), or "false-positive", which was not accompanied by clinical findings. The remaining cases were regarded as "inconclusive".. The one-year cumulative incidences of IA and positive GM tests were 10.1% and 48.1%, respectively. Among 148 episodes of positive GM tests, 97(65.5%), 23(15.5%), and 28(19.0%) were classified as false-positive, true-positive and inconclusive, respectively. In the first episodes of positive GM tests in each patient (false-positive = 67, others = 30), an increase in the GM value in the first two measurements, neutropenia, and use of anti-mold agents at positive GM episode were associated with a significantly lower possibility of false-positive results according to a multivariate analysis.. A false-positive GM test was frequently seen after allogeneic HSCT. An increase in the GM value may increase its positive predictive value.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Incidence; Kinetics; Male; Mannans; Middle Aged; Neutropenia; Retrospective Studies; Risk Factors; Time Factors; Transplantation, Homologous; Young Adult

The diagnosis of invasive aspergillosis remains a challenge. Detection of galactomannan in serum and bronchoalveolar lavage is a useful tool; however due to methodological and economic reasons, the test frequencies of galactomannan assays vary from daily to weekly, which constitute a risk to the patient. In this study, we aimed to evaluate and correlate the performance of the new kit Aspergillus-LFD with the GM-EIA. Aspergillus-LFD kit represents a fast, economical and simple test; showed a good performance and excellent correlation with GM-EIA kit. Given the above, the Aspergillus-LFD is emerging as an alternative to consider in the early diagnosis of invasive aspergillosis.

Topics: Aspergillosis; Aspergillus; Biomarkers; Chile; Chromatography, Affinity; Galactose; Humans; Immunoenzyme Techniques; Mannans; Reagent Kits, Diagnostic; Sensitivity and Specificity; Time Factors

The Platelia Aspergillus Ag assay (Bio-Rad) is designed for detecting Aspergillus galactomannan (GM) and is widely used for diagnosing invasive aspergillosis but is hampered by variable occurrences of unreproducible positive results. Frequency and origin of these unreproducible results have not been formally studied.. Different technicians simultaneously performed four tests on 550 consecutive sera from adult patients (Test#1-Test#2 for extraction#1 and Test#3-Test#4 for extraction#2). The samples were classified as confirmed negative [all tests with GM optical density index (GM-ODI) <0.5], confirmed positive (all tests with GM-ODI ≥0.5), extraction unreproducible positive (Test#1 and Test#2 ODIs ≥0.5, and Test#3 and Test#4 GM-ODIs <0.5, or conversely), and ELISA unreproducible positive (only one test with GM-ODI ≥0.5). The samples with positive and negative GM-ODIs within the assay coefficient of variation values were classified as non-conclusive. Four similar additional tests were performed after ≤72h storage at 4 °C and a new GM test after 8 months at -20 °C.. Five-hundred-twenty sera (94.5%) were confirmed negative, 15 (2.7%) confirmed positive, 4 (0.7%) extraction unreproducible positive, 6 (1.1%) ELISA unreproducible positive, and 5 (0.9%) non-conclusive. Upon retesting, the unreproducible positive results turned negative except for one which turned non-conclusive. The confirmed positive and non-conclusive had similar GM-ODIs (p>0.4) upon retesting after storage ≤72h at 4 °C (n = 20) or eight months at -20 °C (n = 17).. Operational unreproducible positives represent 33% of the GM-positive results and a second sample evaluation appears mandatory to avoid useless investigations or treatments. When operational artifacts are excluded, GM remains stable at standard storage conditions.

Topics: Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Galactose; Humans; Mannans; Reproducibility of Results

Patients with persistent or recurrent neutropenic fevers at risk of invasive fungal disease (IFD) are treated empirically with antifungal therapy (AFT). Early treatment using a diagnostic-driven (DD) strategy may reduce clinical and economic burdens. We compared costs and outcomes of both strategies from a UK perspective.. An empirical strategy with conventional amphotericin B deoxycholate (C-AmB), liposomal amphotericin B (L-AmB), or caspofungin was compared with a DD strategy (initiated based on positive ELISA results for galactomannan antigen) and/or positive results for Aspergillus species on polymerase chain reaction assay) using C-AmB, voriconazole, or L-AmB in a decision-analytic model. Rates of IFD incidence, overall mortality, and IFD-related mortality in adults expected to be neutropenic for ≥10 days were obtained. The empirical strategy was assumed to identify 30% of IFD and targeted AFT to improve survival by a hazard ratio of 0.589. AFT-specific adverse events were obtained from a summary of product characteristics. Resource use was obtained, and costs were estimated by using standard UK costing sources. All costs are presented in 2012 British pounds sterling.. Total costs were 32% lower for the DD strategy (£1561.29) versus the empirical strategy (£2301.93) due to a reduced incidence of adverse events and decreased use of AFT. Administration of AFT was reduced by 41% (DD strategy, 74 of 1000; empirical strategy, 125 of 1000), with similar survival rates.. This study suggests that a DD strategy is likely to be cost-saving versus empirical treatment for immunocompromised patients with persistent or recurrent neutropenic fevers.

Topics: Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus; Caspofungin; Cost Savings; Decision Trees; Deoxycholic Acid; Drug Combinations; Echinocandins; Febrile Neutropenia; Galactose; Health Care Costs; Health Resources; Humans; Immunocompromised Host; Lipopeptides; Mannans; Survival Rate; Voriconazole

Recent products of piperacillin/tazobactam (PTZ) from the original manufacturer, previously considered a major cause of galactomannan (GM) false-positivity, are reported not to be related to it. However, data regarding generic PTZ are limited and controversial. To evaluate the effect of generic PTZ on GM false-positivity in Korea, we performed a case-control study in adult patients with cancer. A case-control study was designed. Electronic medical records of cancer patients who were admitted and tested for serum GM between March and June 2014 at a tertiary care university hospital were reviewed. During the study period, a single generic PTZ (C manufacturer, Korea) was used. Patients who received PTZ within 24 h prior to serum GM testing were enrolled. Age- and GM test date-matched non-PTZ patients were selected as controls. A total of 110 patients received PTZ within 24 h prior to serum GM testing during the study period. The GM optical density index (ODI) of the PTZ group did not vary significantly from that of the control group (p = 0.251). The percentage of false-positive patients in the PTZ group was also similar to that of the control group (p = 0.538). There was no statistical relationship between GM ODI titer and time interval from PTZ administration (p = 0.095) or cumulative PTZ dose (p = 0.416). In a case-control study that evaluated 220 patients, a generic PTZ in Korea was not related to GM false-positivity.

Topics: Adult; Aged; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Case-Control Studies; False Positive Reactions; Female; Galactose; Humans; Male; Mannans; Middle Aged; Neoplasms; Penicillanic Acid; Piperacillin; Retrospective Studies; Tazobactam; Time Factors

Central nervous system (CNS) aspergillosis with stroke has a high mortality and poor prognosis generally. We report a 78-years-old woman with diabetes mellitus, who developed invasive paranasal sinus aspergillosis with the orbital apex syndrome on the right side and cerebral infarction caused by intracranial occlusion of the right internal carotid artery. Based on the presence of a mass lesion in the ethmoid sinus extending to the orbital apex on the right side with cranial CT, the mass lesion was surgically removed and the pathological examination of the surgical specimen revealed aspergillus mold. Immediately after surgery, we initiated treatment with voriconazole 200 mg × 2/day intravenously for 38 days, and then via feeding tube for 86 days until the galactomannan-aspergillus antigen level in the cerebrospinal fluid became negative at 132 days. She is alive now for almost two years without relapse of aspergillosis. There is no definitive guideline for management of patients with CNS aspergillosis concerning the length of drug treatment and the method for monitoring the response for treatment. We believe that measurement of the galactomannan-aspergillus antigen level in the cerebrospinal fluid might be a useful way of monitoring the efficacy of treatment for CNS aspergillosis.

Topics: Aged; Antifungal Agents; Antigens, Fungal; Arterial Occlusive Diseases; Aspergillosis; Biomarkers; Carotid Artery, Internal; Central Nervous System Diseases; Diabetes Complications; Diagnostic Imaging; Female; Galactose; Humans; Infusions, Intravenous; Mannans; Paranasal Sinus Diseases; Stroke; Survival; Time Factors; Voriconazole

To evaluate the clinical value of serum 1,3-beta-D-glucan (BG) and galactomannan (GM) detection for early diagnosis of invasive aspergillosis (IA) in patients after renal transplantation.. Blood samples collected from 69 renal transplant recipients were divided into diagnosis group, clinical diagnosis group, suspected diagnosis group, and non-infected group for detection of serum BG and GM.. The mean serum levels of BG in the diagnosis group, clinical diagnosis group, and suspected diagnosis group were significantly higher than that in non-infected group (P<0.05). The sensitivity, specificity, and positive and negative predictive values of BG was 69.49%, 70%, 93.18% and 35.71% for IA diagnosis, respectively. The serum levels of GM in the 3 diagnosis groups were also significantly higher than that in the non-infected group (P<0.05) with the sensitivity, specificity, and positive and negative predictive values of 84.75%, 90%, 96.15% and 52.63% for IA diagnosis, respectively.. Increased serum BG and GM levels can serve as the evidence for early diagnosis of IA with a high diagnostic sensitivity and specificity in renal transplant recipients.

Topics: Aspergillosis; beta-Glucans; Early Diagnosis; Galactose; Humans; Kidney Transplantation; Mannans; Sensitivity and Specificity

Aspergillus infections are major causes of morbidity and mortality among immunocompromised patients. This study was designed to investigate the galactomannan assay optical density (OD) indices relative to the culture results in bronchoscopic samples obtained from neutropenic and non-neutropenic patients. Galactomannan OD indices from 1427 samples from 2005 to 2012, which were sent from 839 patients and were composed of bronchial lavage (BL = 727) and bronchoalveolar lavage fluids (BAL = 700), were retrospectively analysed. The recovery rates of Aspergillus species from these specimens were 9.4% from the combined patient group and 13.3% from the neutropenic group. Aspergillus fumigatus complex was the most frequently isolated species. The mean and median OD indices of the positive and negative culture samples are approximately 5 and 1, respectively, and 91% of all culture-positive samples have ≥1 OD index value. The receiver-operating characteristics curve analysis demonstrated that the feasibility of the Aspergillus galactomannan assay and Aspergillus galactomannan test has superior accuracy in BAL compared to BL fluids, and the test is not affected by the immune status of the patient. We suggest that the Aspergillus galactomannan test, which uses bronchoscopic material, leads to an earlier diagnosis and if the OD index is found ≥1, fungal growth can be expected.

Topics: Aspergillosis; Aspergillus fumigatus; Bronchoalveolar Lavage Fluid; Galactose; Hematologic Neoplasms; Humans; Lung Diseases, Fungal; Mannans; Neutropenia; Retrospective Studies; Specimen Handling

Severe alcoholic hepatitis (AH) has a poor short-term prognosis. Although infections are frequent complications of AH, the incidence of invasive aspergillosis (IA) and its impact on outcome remain unknown.. We prospectively followed 94 biopsy-proven severe AH episodes for 3 months. We retrospectively reviewed our diagnosis of IA based on EORTC/MSG and AspICU criteria, except for host factors.. Fifteen IA (6 proven, 8 probable, and 1 possible) were diagnosed after a median delay of 26 days following diagnosis of AH. The sites of infection were the lungs (n=11) and central nervous system (n=2), while IA was disseminated in 2 cases. Baseline MELD score ≥24 and ICU admission were independent risk factors for IA. Thirteen IA occurred in the context of corticosteroids, and 2 had received no specific treatment for AH. Non-response to corticosteroids at day 7 was not a risk factor for IA, but IA was associated with absence of liver improvement at day 28. Despite antifungal treatment, 3-month transplant-free survival of patients with IA was 0% compared to 53% in those without IA. IA, Lille score ≥0.45, and overt encephalopathy were independent predictors of transplant-free mortality.. IA is a frequent complication of severe AH and carries a very high risk of mortality. Systematic screening for IA should be recommended in these patients. Further studies are needed to identify high-risk populations requiring antifungal prophylactic treatment.

Topics: Adrenal Cortex Hormones; Adult; Aged; Antifungal Agents; Aspergillosis; Cohort Studies; Female; Galactose; Hepatitis, Alcoholic; Humans; Kaplan-Meier Estimate; Male; Mannans; Middle Aged; Prognosis; Prospective Studies; Retrospective Studies; Risk Factors; Young Adult

Invasive aspergillosis is a leading cause of morbidity and mortality in immunocompromised patients, particularly in individuals with haematological malignancy and in haematopoietic stem cell transplant recipients. Nowadays, the galactomannan (GM) assay has been widely used as an indication of invasive aspergillosis, even though the test is known to generate false-positive results. The aim of this study was to compare the performance of GM and real-time PCR (qPCR) to detected Aspergillus in blood samples obtained from high-risk haematological patients. Haematological patients were screened twice weekly with GM testing, which was performed by the Platelia ELISA kit. An additional sample of whole blood (4 ml) was obtained for the purpose of qPCR testing. Sixty-four samples from 12 patients with haematopoietic stem cell transplant or haematological malignancy were studied. The overall accordance between GM and qPCR tests was 96.9 % (62 samples). Only two samples showed contradictory results, with positive GM test and negative real-time PCR results. Based on the high concordance between GM and qPCR in terms of negative results, the main utility of qPCR could be in the confirmation of positive results seen with GM testing.

Topics: Adult; Aspergillosis; Aspergillus; DNA, Fungal; False Positive Reactions; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Male; Mannans; Mass Screening; Middle Aged; Pilot Projects; Real-Time Polymerase Chain Reaction; Young Adult

The aim of the study was to compare the galactomannan antigen (GA) and molecular biology (PCRrt) tests with the culture in the diagnosis of invasive aspergillosis (IA).. Four hundred and seventy two samples were analyzed: 388 respiratory and 84 serum samples from 271 patients. Culture and GA were evaluated in the respiratory samples and GA in the serum samples. PCR was used when discrepancies were observed among culture and GA tests.. The detection of GA in serum was positive in 22 (of 84), 21 had the test positive respiratory sample. Of the 62 sera with negative GA, 45 were also negative respiratory specimens. The culture was positive in 37 of which were positive for GA. Comparing culture with AG, it showed PPV=23%, NPV=100%, S=100% and E=52%. The PCR showed respect to culture: PPV=69%, NPV=89%, S=64% and E=82%. In sera were found in 60% discrepancies between PCRrt and GA.. We consider useful the GA detection in serum combined with culture and GA in respiratory samples, for diagnosis of AI. PCR requires further studies for standardization and set breakpoints.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Aspergillus; Child; Child, Preschool; Female; Galactose; Humans; Infant; Male; Mannans; Real-Time Polymerase Chain Reaction; Sputum

Piperacillin-tazobactam (PTZ) is known to cause false-positive results in the Platelia Aspergillus enzyme-linked immunoassay (EIA), due to contamination with galactomannan (GM). We tested 32 lots of PTZ and 27 serum specimens from patients receiving PTZ. GM was not detected in the lots of PTZ; one serum specimen (3.7%) was positive. PTZ formulations commonly used in the United States today appear to be a rare cause for false-positive GM results.

Topics: Aged, 80 and over; Anti-Bacterial Agents; Aspergillosis; Aspergillus; Diagnostic Tests, Routine; Drug Contamination; False Positive Reactions; Female; Galactose; Humans; Immunoenzyme Techniques; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Serum; United States

Research to develop and validate novel methods for diagnosis of aspergillosis based on detection of galactomannan requires the use of clinical specimens that have been stored frozen. Data indicating that galactomannan remains stable when frozen are scant. The objective of this study was to determine the stability of galactomannan in clinical specimens stored at -20 °C that were positive in the Platelia Aspergillus enzyme immunoassay when initially tested. Prospective real-time testing of serum and bronchoalveolar lavage (BAL) fluid pools from positive and negative patient specimens showed no decline in galactomannan index (GMI) over 11 months at -20 °C and no development of positive reactions in the negative-control pool. Retrospective testing of positive specimens that had been stored at -20 °C for 5 years showed that 28 of 30 serum (n = 15) or BAL (n = 15) specimens remained positive. These findings support the use of frozen serum or BAL specimens stored for at least 5 years in evaluation of diagnostic tests based on detection of galactomannan.

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Freezing; Galactose; Humans; Immunoenzyme Techniques; Mannans; Serum; Specimen Handling; Time Factors

Invasive fungal infections remain major causes of infection-related mortality in hematopoietic stem cell transplantation (HSCT) patients. Mixed infections and multiple organ involvement have been reported in these patients. Here, we report a case of mixed Aspergillus and Mucorales infection involving the lungs, brain, spleen and bone in a HSCT patient with relapsed acute myeloid leukemia, who finally improved with triple antifungal therapy and neurosurgical evacuation of brain abscesses. She was put on lifelong secondary prophylaxis with posaconazole with excellent compliance and no sign of toxicity despite over 10 years of drug administration. Serial galactomannan measurements and positron emission tomography/computed tomography were used and were helpful for disease activity monitoring. This is an instructive case of long-term survival after a severe combined mould infection.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Bone Marrow; Brain; Chemoprevention; Coinfection; Drug Monitoring; Female; Galactose; Humans; Lung; Mannans; Mucorales; Mucormycosis; Spleen; Survivors; Tomography, X-Ray Computed; Treatment Outcome; Young Adult

Invasive aspergillosis is a life-threatening infection in immunocompromised patients, and treating these infections at an early stage is often crucial for a favorable outcome. Early diagnosis, however, remains challenging. We performed a retrospective comparison of three methods: real-time quantitative PCR (qPCR), nucleic acid sequence-based amplification (NASBA), and galactomannan enzyme-linked immunosorbent assay (GM-ELISA); these detect circulating Aspergillus DNA, RNA, and galactomannan, respectively. Blood samples from 80 patients at high risk for invasive aspergillosis were tested by each assay. The sensitivity of NASBA, qPCR, and GM-ELISA was 76.47% (95% CI, 58.4-88.6%), 67.65% (95% CI, 49.4-82.0%), and 52.94% (95% CI, 35.4-69.8%), respectively, and the specificity was 80.43% (95% CI, 65.6-90.1%), 89.13% (95% CI, 75.6-95.9%), and 80.43% (95% CI, 65.6-90.1%), respectively. We also evaluated the efficiency of the three tests in various combinations. Perfect specificity (100%; 95% CI, 90.4-100%) and perfect positive predictive value (100%; 95% CI, 77.1-100%) were achieved by combining NASBA and qPCR testing in series. Testing with both NASBA and qPCR in parallel was the most sensitive and had the highest Youden index. Our data support the great potential of NASBA and qPCR, singly or in combination, for diagnosis of invasive aspergillosis in high-risk populations.

Topics: Aged; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Real-Time Polymerase Chain Reaction; Reproducibility of Results; Retrospective Studies; ROC Curve; Self-Sustained Sequence Replication

There is a practical need to investigate the performance of the serum galactomannan (GM) assay in hematology patients with a potentially low pretest risk of invasive aspergillosis following effective antimold prophylaxis.. We present a 4-year study with 262 unselected consecutive high-risk episodes, prospectively managed with posaconazole primary prophylaxis and a uniform diagnostic algorithm, including biweekly serum GM quantification for early detection of invasive aspergillosis.. A total of 2972 serum GM tests were performed (median, 11 per episode [range, 3-30]); the vast majority were negative (96.7% of tests and 83.6% of episodes). The incidence of breakthrough invasive aspergillosis was 1.9% (5/262), all with true-positive GM test results. Our study identified 30 false-positive GM evaluable episodes (85.7%; 13.8% of all evaluable episodes), validating with real-life data the low positive predictive value of the assay in this setting (12%). In 26 of these 30 episodes (86.7%), the false-positive result(s) occurred in tests performed as preemptive surveillance only. Conversely, in evaluable cases with positive GM tests and a clinical suspicion of invasive fungal disease, the performance of diagnostic-driven GM tests improved, with a positive predictive value of 89.6%.. The low pretest risk of invasive aspergillosis in the context of effective antimold prophylaxis renders serum GM surveillance of asymptomatic patients unreliable, as all results would be either negative or false positive. The test remains useful to diagnose patients with a clinical suspicion of invasive fungal disease, calling for a more efficient copositioning of effective prophylaxis and GM testing in this clinical setting.

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Galactose; Humans; Mannans; Triazoles

Aspergillus fumigatus remains a major respiratory pathogen in birds and treatment is still difficult. We challenged different groups of few-day-old turkeys via intratracheal aerosolisation with increasing concentrations (10(5) up to 10(8)) of conidia using a MicroSprayer(®) device. The fungal burden was assessed by real-time PCR, galactomannan dosage, CFU counting and histopathological evaluation in order to provide a comparison of these results within each inoculum groups. Significant mortality, occurring in the first 96h after inoculation, was only observed at the highest inoculum dose. Culture counts, GM index and qPCR results on the one hand and inoculum size on the other hand appeared to be clearly correlated. The mean fungal burden detected by qPCR was 1.3log10 units higher than the mean values obtained by CFU measurement. The new model and the markers will be used to evaluate the efficacy of antifungal treatments that could be used in poultry farms.

Topics: Aerosols; Animals; Aspergillosis; Aspergillus fumigatus; Colony Count, Microbial; Galactose; Mannans; Poultry Diseases; Real-Time Polymerase Chain Reaction; Spores, Fungal; Survival Analysis; Turkeys

To investigate the invasive aspergillosis (IA) status in a surgical intensive care unit (SICU).. The clinical data including general state, operation, pathogenic microorganisms, infection position, clinical situation, treatment and prognosis of patients with IA admitted to the SICU of Peking University First Hospital from January 2004 to December 2013 were retrospectively analyzed.. 8 220 patients admitted to the SICU of Peking University First Hospital from January 2004 to December 2013 were enrolled. Of 8 220 patients, there were 29 cases experienced IA, with an incidence of 0.35%, and the incidence of hospital onset of IA infection was 0.27% (22/8 220). The incidence of hospital onset of IA infection was accounted for 6.98% (22/315) of the incidence of hospital onset of infection in SICU in the same period. Compared with 2004-2008, in 2009-2013, the incidence of hospital onset of infection was significantly decreased [3.19% (137/4 293) vs. 4.53% (178/3 927), χ² = 10.020, P=0.002], while the incidence of IA [0.56% (24/4 293) vs. 0.13% (5/3 927), χ² = 10.874, P=0.001], the incidence of hospital onset of IA infection [0.40% (17/4 293) vs. 0.13% (5/3 927), χ² = 5.556, P=0.019], and the percentage of incidence of hospital onset of IA infection according to the incidence of hospital onset of infection were increased by 5 years [12.40% (17/137) vs. 2.81% (5/178), χ² = 10.982, P=0.001]. Of 29 patients with IA, 25 cases had occurred after operation, and the majority of them were from the Department of General Surgery (13 cases), and followed by post-renal transplantation (6 cases) and post-thoracic surgery (3 cases). In the total submission of 155 specimens from 29 patients with IA, there were 17 strains isolated aspergillosis, among which included 2 strains of Aspergillus fumigatus, and 15 other un-subgrouped strains. The most common infection site was lower respiratory tract (23 cases, 79.31%). Sixteen patients were found with positive galactomannan (GM) test. In all the risk factors contributing to IA, the ratio of the long-term usage of broad-spectrum antibiotics over 4 days was the highest [36.25% (29/80)], which followed by the long-term use of hormone [18.75% (15/80)], complicated with acute kidney injury [18.75% (15/80)], liver injury [13.75% (11/80)], the long-term use of immunosuppressive orally [7.50% (6/18)], and complicated with neutropenia [5.00% (4/80)]. In 29 patients with IA, there were 28 patients received anti-fungal treatment except for 1 patient without treatment, and those were single use of itraconazole in 1 case, single use of echinocandins in 3 cases, single use of liposomal amphotericin B in 5 cases, 8 cases with voriconazole, single use of liposomal amphotericin B or echinocandins then replaced by voriconazole in 8 cases, and 3 cases of echinocandins therapy combined with voriconazole. Seventeen of 29 patients di. IA is an uncommon but increasing postoperative complication of patients in SICU in recent 5 years. The most common sites of IA were lower respiratory tract. The mortality of IA is very high. So clinicians should pay more attention to the careful monitor for IA.

Topics: Amphotericin B; Aspergillosis; Critical Care; Galactose; Humans; Incidence; Intensive Care Units; Mannans; Prognosis; Retrospective Studies; Risk Factors

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Clinical Laboratory Techniques; Galactose; Humans; Mannans; Serum; Specimen Handling; Spectrophotometry

Samples from patients at high risk for invasive aspergillosis (IA) were prospectively collected and analyzed for the presence of molecular markers of fungal infection. Serum specimens were screened for galactomannan and Aspergillus DNA, and whole-blood specimens were screened only for Aspergillus DNA. Fungal infections were categorized according to the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group, National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) criteria. Forty-seven cases (proven and probable IA) and 31 controls (no evidence of IA) were selected retrospectively for this case-control study, comprising 803 samples, in order to determine the performance of whole-blood PCR, serum PCR, and serum galactomannan testing. Although no single assay was able to detect every case of IA, a combination of different assays provided the best performance. There was no significant difference between the use of whole-blood and serum specimens for PCR-based diagnosis of IA, but there was a trend for whole blood to be more sensitive (85% versus 79%) and to yield an earlier positive result (36 days versus 15 days) than for serum. However, DNA extraction from serum specimens is easier and faster than that from whole-blood specimens, and it allows the same specimen to be used for both galactomannan and PCR assays. In conclusion, the appropriate sample type for DNA extraction should be determined by the local requirements and the technical platforms available at each individual center. A combination of biomarker tests offered the best diagnostic utility for detecting IA.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; Biomarkers; Case-Control Studies; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Retrospective Studies; Young Adult

We report a case of Listeria monocytogenes bacteremia in a leukemic patient having a positive assay for aspergillus galactomannan (GM), although no evidence of aspergillosis was found. Supernatant obtained from L. monocytogenes strain suspension was reactive with GM-assay. L. monocytogenes produces a soluble antigen that is cross-reactive with Aspergillus GM.

Topics: Anti-Bacterial Agents; Antifungal Agents; Antigens, Fungal; Antineoplastic Agents; Aspergillosis; Aspergillus; Bacteremia; Cross Reactions; Galactose; Humans; Leukemia; Listeria monocytogenes; Male; Mannans; Middle Aged

Invasive aspergillosis (IA) is a leading complication of intensive treatment for haematological malignancies. Earlier diagnosis should facilitate effective antifungal therapy and prevent progression to invasive disease, which is often lethal. Polymerase chain reaction (PCR) assays, targeting the 28S and ITS ribosomal gene regions respectively, were evaluated for early detection of Aspergillus DNA and for diagnostic utility in patients receiving treatment in two busy haematopoietic stem cell transplant centres. Patients undergoing intensive chemotherapy, autologous or allogeneic transplant were eligible for inclusion in the study. EDTA blood and serum samples for circulating Aspergillus biomarkers, including galactomannan (GM), were collected twice-weekly on a prospective basis from all study patients who were categorized according to international consensus criteria for defining invasive fungal disease (IFD). Of 278 patients recruited there were 44 probable IA cases and only one proven case. Moderate sensitivity and specificity, poor positive predictive value (50-80%), but good negative predictive value (>80-90%) were common to both PCR assays. Overall biomarker performance could be improved by combining positive results of either PCR assay with GM taken within a 12-d period. The addition of PCR to GM monitoring in high-risk patients with haematological malignancies provides greater diagnostic accuracy in invasive aspergillosis.

Topics: Aspergillosis; Aspergillus; DNA, Ribosomal Spacer; Galactose; Hematologic Neoplasms; Humans; Incidence; Mannans; Prevalence; Real-Time Polymerase Chain Reaction; RNA, Ribosomal, 28S; Sensitivity and Specificity; Time Factors; Tomography, X-Ray Computed

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Female; Galactose; Humans; Leukemia, Lymphoid; Male; Mannans; Polymerase Chain Reaction

The detection of galactomannan in serum is a cornerstone for the diagnosis of invasive fungal disease (IFD). Because a delay in treatment initiation is associated with a poor outcome, the results have to be available promptly. However, due to methodological and economic reasons, the test frequencies of the commonly used galactomannan assays vary between daily to weekly, meaning that results may be available too late to be clinically useful. The novel Aspergillus lateral-flow device (Aspergillus-LFD) is a rapid test that may overcome these limitations.. We compared the diagnostic performance of the Aspergillus-LFD and the Platelia® Aspergillus EIA (GM-EIA) in serum from 101 patients during and after allogeneic haematopoietic stem cell transplantation (HSCT). Clinical data and sera were collected prospectively and patients classified according to the European Organisation for Research and Treatment of Cancer (EORTC)/Mycoses Study Group (MSG) 2008 guidelines.. By the end of hospitalisation, one proven, nine probable and 20 possible cases of IFD were identified. Depending on the number of positive serum samples required for test positivity, the sensitivities, specificities and diagnostic odds ratios in patients with proven and probable IFD were as follows. One positive serum required: Aspergillus-LFD 40.0 %, 86.8 % and 3.03; GM-EIA 40.0 %, 89.0 % and 3.64. Two positive sera required: Aspergillus-LFD 20.0 %, 97.8 % and 11.13; GM-EIA 30.0 %, 98.9 % and 38.57. Although the GM-EIA was positive in a higher percentage of samples, this did not result in an earlier diagnosis.. If used as a screening test (one positive serum required for test positivity) or to rule out IFD, the Aspergillus-LFD has shown a comparable diagnostic performance to the GM-EIA. However, if the results have to be confirmed by a second positive serum, the GM-EIA exhibited superior sensitivity. In terms of practicability, the Aspergillus-LFD has demonstrated to be a quick (15 min) and easy-to-use test for single-patient detection of Aspergillus antigens.

Topics: Adult; Aged; Aspergillosis; Aspergillus; Biomarkers; Chromatography, Affinity; False Positive Reactions; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunoenzyme Techniques; Male; Mannans; Middle Aged; Reagent Kits, Diagnostic; Young Adult

Patients with cystic fibrosis (CF) demonstrate a wide range of hypersensitivity responses to Aspergillus, beyond allergic bronchopulmonary aspergillosis, which require classification.. This study integrated 2 new methods of Aspergillus detection-sputum galactomannan (GM) and real-time PCR-alongside established serologic markers, to reclassify aspergillosis in CF.. A total of 146 adult patients with CF had serologic tests (ImmunoCap total IgE, specific Aspergillus fumigatus IgE, and specific A fumigatus IgG), sputum real-time Aspergillus PCR, and sputum GM. Patients were classified by using latent class analysis.. Both RT-PCR and GM were more sensitive than culture in detecting Aspergillus in sputum (culture 37%, RT-PCR 74%, and GM 46%). Intraassay and interassay reproducibility of PCR and GM was excellent. Latent class analysis of triazole-naive patients identified a nondiseased group and 3 disease classes: class 1 (n = 49, 37.7%) represented patients with or without positive RT-PCR but no immunologic response to A fumigatus and negative GM (nondiseased); class 2 (n = 23, 17.7%) represented patients with positive RT-PCR, elevated total and specific A fumigatus IgE/IgG, and positive GM (serologic allergic bronchopulmonary aspergillosis); class 3 (n = 19, 14.6%) represented patients with or without positive RT-PCR, elevated A fumigatus IgE (not IgG), and negative GM (Aspergillus sensitized); and class 4 (n = 39, 30%) represented patients with positive RT-PCR, elevated A fumigatus IgG (not IgE), and positive GM (Aspergillus bronchitis).. Three distinct classes of aspergillosis in CF were identified by latent class analysis by using serologic, RT-PCR, and GM data. This novel classification will facilitate improved phenotyping, pathogenesis studies, and management evaluations.

Topics: Adult; Allergens; Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Cystic Fibrosis; Female; Galactose; Humans; Immunoglobulin E; Immunoglobulin G; Male; Mannans; Prospective Studies; Real-Time Polymerase Chain Reaction; Skin Tests; Sputum; Young Adult

To explore the relationship between the optical density index of serum aspergillus galactomannan (GM) assay and invasive aspergillosis (IA).. From Jan 2008 to Dec 2011, 825 hematological diseases patients with neutrophil count <0.5×10⁹/L⁹ by continuous blood count tests were admitted into our hospital. The optical density index of GM assay was ≥0.5 at least once. Of 825 patients, 247 cases were manifested as fever during hospitalization. The optical density index of GM antigen was detected by enzyme-linked immunosorbent assay, and the sensitivity and specificity of optical density ranged in 0.5-1.5.. In this study, the sensitivity and specificity of GM assay with continuous twice samples (73% and 93%, respectively) were higher than single sample (66% and 80%, respectively) when optical density index ≥1.0. 69 cases were diagnosed as proven IA with the incidence rate of 8.36%.. The cut-off level for serum GM antigen assay should be decided as optical density index in two continuous samples of ≥1.0.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Diseases; Humans; Male; Mannans; Middle Aged; Sensitivity and Specificity; Young Adult

Galactomannan (GM) was recently included in consensus guidelines as an indirect mycological criterion for the diagnosis of invasive aspergillosis. Currently, there is an enzyme immunoassay available to detect GM in biological samples, the Platelia™ Aspergillus EIA. In this study, the reproducibility of positive results obtained using this assay was evaluated using serum samples from neutropenic patients. A trend toward lower values was observed, and 55 %(27/49) of positive results were negative after retesting. A low reproducibility of positive results for the detection of GM in serum was observed.

Topics: Aspergillosis; Galactose; Humans; Immunoenzyme Techniques; Mannans; Reproducibility of Results; Serum

Measurement of serum galactomannan (GM), a polysaccharide fungal cell-wall component, is a non-invasive test for early diagnosis of invasive aspergillosis in humans. Feline upper respiratory tract (URT) aspergillosis is an emerging infectious disease in cats. Diagnosis requires biopsy for procurement of tissue specimens for cytological or histological detection of fungal hyphae and for fungal culture. The aim of this study was to evaluate serum GM measurement as a non-invasive diagnostic test for URT aspergillosis in cats. A one-stage, immunoenzymatic sandwich ELISA was used to detect serum GM in 4 groups of cats; Group 1 (URT aspergillosis) - confirmed URT aspergillosis (n=13, sinonasal aspergillosis (SNA) n=6 and sino-orbital aspergillosis (SOA) n=7), Group 2 (URT other) - other URT diseases (n=15), Group 3 (β-lactam) - cats treated with β-lactam antibiotics for non-respiratory tract disease (n=14), Group 4a - healthy young cats (≤ 1 y of age, n=28), Group 4b - healthy adult cats (>1 y of age, n=16). One cat with SNA and two cats with SOA caused by an Aspergillus fumigatus-mimetic species, tested positive for serum GM. For a cut-off optical density index of 1.5, the overall sensitivity and specificity of the assay was 23% and 78% respectively. False positive results occurred in 29% of cats in Group 3 and 32% of cats in Group 4a. Specificity increased to 90% when Groups 3 and 4a were excluded from the analysis. Overall, serum GM measurement has a poor sensitivity but is a moderately specific, non-invasive screening test to rule out infection in patients with suspected feline upper respiratory tract aspergillosis.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Cat Diseases; Cats; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Lung Diseases, Fungal; Male; Mannans; Sensitivity and Specificity

Invasive aspergillosis (IA) is a major cause of morbidity and mortality in immunocompromised children. We investigated the usefulness of an Aspergillus galactomannan (GM) antigen assay as a diagnostic tool for IA in pediatric cancer patients and hematopoietic cell transplantation (HCT) recipients.. The GM antigen assay results were analyzed in 749 blood samples from 99 patients. A GM index (GMI) greater than or equal to 0.5 on at least two separate occasions was considered positive. A review of the clinical data was performed for subjects with proven or probable IA.. Twenty-one of 23 patients with proven or probable IA had positive GM antigen test results (91.3% sensitivity, 95% CI 71.9-98.9; 81.7% specificity, 95% CI 69.6-90.5; P < 0.0001). The false-positive rate was 18.3%. Being younger than 3 years of age, having a solid tumor, and receiving HCT within 4 weeks of the test were statistically significant factors for causing false-positive results (P < 0.05). Among the 23 patients with IA (six proven, 17 probable), 16 (69.6%) had hematological malignancies, five (22.7%) had solid tumors, and two (8.7%) had primary immunodeficiency. Nineteen patients (82.6%) received HCT. The most common clinical site of IA was the lungs (91.3%), and consolidation was the most frequent finding in chest CT scans (36.8%). The mortality at 12 weeks was 43.5%.. Having a positive GM assay at least twice is useful in diagnosing IA in pediatric patients with cancer and HCT recipients.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Child; Child, Preschool; False Positive Reactions; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Immunoenzyme Techniques; Infant; Male; Mannans; Sensitivity and Specificity

To evaluate risk factors, diagnostic procedures, and treatment outcomes of invasive aspergillosis (IA) in patients with hematological malignancies.. A retrospective analysis of data from proven/probable IA cases that occurred from 2005 to 2009 at 10 hematology centers was performed.. We identified 176 IA cases that mainly occurred in patients with acute leukemias (58.5%), mostly those on induction/re-induction treatments (39.8%). Prolonged neutropenia was the most frequent risk factor for IA (61.4%). The lungs were the most frequently affected site (93.8%) and computed tomography detected abnormalities in all episodes; however, only 53.7% of patients had findings suggestive of IA. Galactomannan (GM) detection in serum or bronchoalveolar lavage fluid (positive in 79.1% and 78.8% of episodes, respectively) played a crucial role in IA diagnosis. Neutrophil count and antifungal prophylaxis did not influence the GM positivity rate, but empirical therapy decreased this rate (in serum). Of the IA cases, 53.2% responded to initial antifungal therapy. The combination of voriconazole and echinocandin, even as initial or salvage therapy, did not perform better than voriconazole monotherapy (p=0.924 for initial therapy and p=0.205 for salvage therapy). Neutrophil recovery had a significant role in the response to initial (but not salvage) antifungal therapy.. Our retrospective analysis identified key diagnostic and treatment characteristics, and this understanding could improve the management of hematological malignancy patients with IA.

Topics: Acute Disease; Adolescent; Adult; Aged; Antifungal Agents; Aspergillosis; Bronchoalveolar Lavage Fluid; Child; Child, Preschool; Czech Republic; Databases, Factual; Echinocandins; Female; Galactose; Humans; Leukemia; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Neutrophils; Pyrimidines; Retrospective Studies; Slovakia; Triazoles; Voriconazole; Young Adult

We have evaluated the in vitro activity of voriconazole against 61 strains of Aspergillus fumigatus by using broth microdilution, disk diffusion, and minimal fungicidal concentration procedures. We observed an excellent correlation between the results obtained with the three methods. Five percent of the strains showed MICs greater than or equal to the epidemiological cutoff value (ECV; 1 μg/ml). To assess whether MICs were predictive of in vivo outcome, we tested the efficacy of voriconazole at 25 mg/kg of body weight daily in an immunosuppressed murine model of disseminated infection using 10 strains representing various patterns of susceptibility to the drug as determined by the in vitro study. Voriconazole prolonged survival and reduced fungal load in the kidneys and brain in those mice infected with strains with MICs of ≤0.25 μg/ml, while in mice infected with strains with MICs of 0.5 to 2 μg/ml, the efficacy was varied and strain dependent and in mice infected with the strain with a MIC of 4 μg/ml, the antifungal did not show efficacy. Voriconazole reduced galactomannan antigenemia against practically all strains with a MIC of <4 μg/ml. Our results demonstrate that some relationship exists between voriconazole MICs and in vivo efficacy; however, further studies testing additional strains are needed to better ascertain which MIC values can predict clinical outcome.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Brain; Drug Resistance, Fungal; Galactose; Immunocompromised Host; Kidney; Male; Mannans; Mice; Microbial Sensitivity Tests; Predictive Value of Tests; Pyrimidines; Triazoles; Voriconazole

We evaluated the efficacy of voriconazole against nine strains of Aspergillus terreus with different MICs (0.12 to 4 μg/ml) by using a murine model. Markers of efficacy included survival, tissue burden, galactomannan antigenemia, and drug serum levels. Voriconazole was especially effective in prolonging survival and reducing the fungal load in infections by strains that showed MICs that were less than or equal to the epidemiological cutoff value (1 μg/ml). In vitro data might be useful for predicting the outcome of A. terreus infections.

Topics: Amphotericin B; Animals; Antifungal Agents; Aspergillosis; Aspergillus; Drug Resistance, Fungal; Galactose; Male; Mannans; Mice; Microbial Sensitivity Tests; Prognosis; Pyrimidines; Survival Analysis; Triazoles; Voriconazole

Diagnosis of invasive aspergillosis for patients with high risk of infection is based on the monitoring of Aspergillus antigenemia assessed by the detection of galactomannan in serum by a sandwich-type ELISA (Biorad(®)). The validation of the method was displayed according to the guide COFRAC SH GTA 04. The internal quality control system settled, involves two quality control samples made of pools of sera (negative and positive). The repeatability of the measurements, as estimated by the coefficients of variation (CV), obtained by two different technicians was found from 9 to 13.7% for the positive control. The CV of the negative control, for which the provider indicates it is not useful in the analytical process, was found from 7.1 to 30%. In our experience it could be an indicator of environmental contamination. The evaluation of the intermediary fidelity was 15.7% for the positive control and 22.5% for the negative one. In the lack of reference material (International Standard) and recommendation from scientific societies, performances obtained will be discussed according to the results reported in the technical form of the supplier and those obtained by 39 laboratories participating in the only available external quality assessment program organized in France by ProBioQual(®) where the CV of reproducibility are 44.7% of unit (mean index 0.131) for the negative control and 18% (mean index 1.089) for the positive one in 2011.

Topics: Accreditation; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Fungemia; Galactose; Humans; Laboratory Proficiency Testing; Mannans; Reproducibility of Results

Diagnostic galactomannan (GM) enzyme immunoassay (EIA) testing is formally validated only for serum, though in practice, plasma is occasionally tested. It is assumed, but not confirmed, that results will be comparable to those for serum. GM EIA when testing plasma was evaluated, providing sensitivity (85.7%) and specificity (85.4%) comparable to those for serum. Plasma index values were higher than those for serum; if plasma GM EIA were used to define probable cases, four additional cases would have been diagnosed.

Topics: Antigens, Fungal; Aspergillosis; Clinical Laboratory Techniques; Galactose; Humans; Immunoenzyme Techniques; Mannans; Plasma; Sensitivity and Specificity; Serum

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Humans; Immunoenzyme Techniques; Laboratories, Hospital; Mannans; Reagent Kits, Diagnostic; Reproducibility of Results

Galactomannan (GM) is a polysaccharide component of the cell wall of Aspergillus spp. and is released into the host's circulation by growing hyphae. GM testing of patients with hematological malignancies has been rarely considered in recent epidemiologic studies of invasive mould infections (IMIs). The aim of the investigation was to analyze the impact of GM testing on the reported prevalence of IMI by comparing detection rates of IMI before and after the introduction of this diagnostic procedure. Prevalence of IMI was assessed by conducting a prospective single-centre study over seven months in 2010. Results obtained were then compared to those obtained with a representative collection of patients assessed by the same investigators at the same institution over seven months in 2007, i.e., prior to the introduction of GM testing. We found that, in general, detection rates of invasive aspergillosis (IA) and invasive mould infections increased significantly after the introduction of GM analysis. This study may therefore indicate that GM testing has a significant impact on the reported prevalence of IMI. Broad usage of such testing in patients with hematological malignancies may be able to produce a realistic picture of IMI rates when current diagnostic criteria are applied.

Topics: Aspergillosis; Aspergillus; Clinical Laboratory Techniques; Female; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Prevalence; Prospective Studies

It is well known that cross reactions with other fungal pathogens including Histoplasma capsulatum can occur with the use of the Platelia™ Aspergillus galactomannan assay. We report two patients with confirmed blastomycosis whose bronchoalveolar lavage (BAL) fluid tested positive for Aspergillus galactomannan despite no evidence of aspergillosis.

Topics: Aged; Aspergillosis; Aspergillus; Blastomycosis; Clinical Laboratory Techniques; Cross Reactions; Galactose; Humans; Immunoenzyme Techniques; Male; Mannans; Mycology

In conventional ΜΙC tests, fungi are exposed to constant drug concentrations, whereas in vivo, fungi are exposed to changing drug concentrations. Therefore, we developed a new in vitro pharmacokinetic/pharmacodynamic model where human plasma pharmacokinetics of standard doses of 1 mg/kg amphotericin B, 4 mg/kg voriconazole, and 1 mg/kg caspofungin were simulated and their pharmacodynamic characteristics were determined against three clinical isolates of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus terreus with identical MICs (1 mg/liter for amphotericin B, 0.5 mg/liter for voriconazole) and minimum effective concentrations (0.5 mg/liter for caspofungin). This new model consists of an internal compartment (a 10-ml dialysis tube made out of a semipermeable cellulose membrane allowing the free diffusion of antifungals but not galactomannan) inoculated with Aspergillus conidia and placed inside an external compartment (a 700-ml glass beaker) whose content is diluted after the addition of antifungal drugs by a peristaltic pump at the same rate as the clearance of the antifungal drugs in human plasma. Fungal growth was assessed by galactomannan production. Despite demonstrating the same MICs, amphotericin B completely inhibited (100%) A. fumigatus but not A. flavus and A. terreus, whose growth was delayed for 7.53 and 22.8 h, respectively. Voriconazole partially inhibited A. fumigatus (49.5%) and Α. flavus (27.9%) but not Α. terreus; it delayed their growth by 3.99 h (A. fumigatus) and 5.37 h (Α. terreus). Caspofungin did not alter galactomannan production in all of the species but A. terreus. The new model simulated human pharmacokinetics of antifungal drugs and revealed important pharmacodynamic differences in their activity.

Topics: Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus; Aspergillus flavus; Aspergillus fumigatus; Caspofungin; Diffusion Chambers, Culture; Echinocandins; Galactose; Humans; Lipopeptides; Mannans; Microbial Sensitivity Tests; Pulsatile Flow; Pyrimidines; Species Specificity; Spores, Fungal; Triazoles; Voriconazole

Invasive aspergillosis is a serious disease, the lethality of which is important among hematology patients. Early diagnosis is crucial for treatment options and the prognosis. Detection of the antigen galactomannan is the most frequently used microbiological tools. But galactomannan detection may be falsely positive, and this false positivity has been associated with piperacillin-tazobactam treatment, the main antibiotic combination used in clinical hematology.. The purpose of our study, carried out from January 2009 to December 2010 at the Versailles hospital on in-patients with hematological disorders, was to evaluate the association between false galactomannan positivity and administration of piperacillin-tazobactam, and to study a possible variability of products issued by three manufacturers.. We noted that 207 patients were included (n=207), accounting for 69 false positive and 138 true negative results. The intrinsic galactomannan values in the study were sensitivity 100%, specificity 68%, positive and negative predictive values respectively 16%, 100%, and a likelihood positive and negative test at respectively 3.12, and 0.. The statistical analysis did not determine any association between false positivity in galactomannan and piperacillin-tazobactam issued by two manufacturers (P=0.87 and P=0.94). But, there was a significant association between false galactomannan positivity and piperacillin-tazobactam issued by the third manufacturer (P=0.02). Four of the 25 batches issued by this manufacturer were tested and negative "in vitro" for galactomannan.. This study results suggest that the association between false galactomannan positivity and piperacillin-tazobactam is not longer systematic, but can still prevail depending on the manufacturers. It also confirmed the positive contribution of testing piperacillin-tazobactam batches "in vitro" before using the antibiotic.

Topics: Anti-Bacterial Agents; Antigens, Fungal; Artifacts; Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Fungemia; Galactose; Humans; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Reagent Kits, Diagnostic; Reproducibility of Results; Retrospective Studies; Sensitivity and Specificity

Nine of 11 hematological patients with disseminated/deep-seated Fusarium infection tested at least twice for Aspergillus galactomannan (GM) had repeated positive results in the absence of Aspergillus isolation in culture. The centrifuged supernatants of 12 Fusarium isolates were tested by a GM enzyme-linked immunosorbent assay (EIA). All the isolates produced positive reactions when tested undiluted. These results show cross-reactivity of Fusarium spp. with Aspergillus GM that may constitute a drawback with respect to the specificity of the Platelia EIA.

Topics: Adult; Aspergillosis; Aspergillus; Child; Child, Preschool; Clinical Laboratory Techniques; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Female; Fusariosis; Fusarium; Galactose; Humans; Male; Mannans; Middle Aged; Young Adult

Topics: Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Biomarkers; Cross Infection; Fatal Outcome; Galactose; Haemophilus influenzae; Hepatitis C, Chronic; Humans; Immunocompromised Host; Liver Cirrhosis; Low Back Pain; Lumbar Vertebrae; Male; Mannans; Middle Aged; Multiple Organ Failure; Opportunistic Infections; Pulmonary Disease, Chronic Obstructive; Pyrimidines; Spondylitis; Triazoles; Voriconazole

Invasive aspergillosis (IA) is an important cause of mortality in critically ill patients, but the diagnosis is difficult as clinical and radiological signs are neither sensitive nor specific. Serum galactomannan (GM) is a useful marker for IA, but exhibits low sensitivity in non-neutropenic patients. In our previous work, strong antibody reactivity to thioredoxin reductase of Aspergillus fumigatus was found in non-neutropenic IA patients.. Using recombinant thioredoxin reductase GliT (TR), an antigenic protein secreted by A. fumigatus, as the coating antigen, an enzyme-linked immunosorbent assay (ELISA) for detecting anti-TR antibodies was developed. The antibody response to TR in IA animal models and 42 non-neutropenic patients with culture- and/or histology-documented IA was investigated. The results showed that anti-TR antibody was detectable in rabbit serum 7-9 days after exposure to the fungus. The sensitivity and specificity of the anti-TR antibody assay in patients were 80.9% and 96%, respectively, while the sensitivity of GM in this group of patients was only 52.3%. The specificity of the assay was confirmed by testing the sera from patients infected with other pathogenic fungal species and bacteria.

Topics: Adult; Aged; Aged, 80 and over; Animals; Antibodies, Fungal; Antibody Specificity; Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Neutropenia; Rabbits; Sensitivity and Specificity; Serologic Tests; Thioredoxin-Disulfide Reductase

The recognition of pathogen-derived structures by C-type lectins and the chemotactic activity mediated by the CCL2/CCR2 axis are critical steps in determining the host immune response to fungi. The present study was designed to investigate whether the presence of single nucleotide polymorphisms (SNPs) within DC-SIGN, Dectin-1, Dectin-2, CCL2 and CCR2 genes influence the risk of developing Invasive Pulmonary Aspergillosis (IPA). Twenty-seven SNPs were selected using a hybrid functional/tagging approach and genotyped in 182 haematological patients, fifty-seven of them diagnosed with proven or probable IPA according to the 2008 EORTC/MSG criteria. Association analysis revealed that carriers of the Dectin-1(rs3901533 T/T) and Dectin-1(rs7309123 G/G) genotypes and DC-SIGN(rs4804800 G), DC-SIGN(rs11465384 T), DC-SIGN(7248637 A) and DC-SIGN(7252229 C) alleles had a significantly increased risk of IPA infection (OR = 5.59 95%CI 1.37-22.77; OR = 4.91 95%CI 1.52-15.89; OR = 2.75 95%CI 1.27-5.95; OR = 2.70 95%CI 1.24-5.90; OR = 2.39 95%CI 1.09-5.22 and OR = 2.05 95%CI 1.00-4.22, respectively). There was also a significantly increased frequency of galactomannan positivity among patients carrying the Dectin-1(rs3901533_T) allele and Dectin-1(rs7309123_G/G) genotype. In addition, healthy individuals with this latter genotype showed a significantly decreased level of Dectin-1 mRNA expression compared to C-allele carriers, suggesting a role of the Dectin-1(rs7309123) polymorphism in determining the levels of Dectin-1 and, consequently, the level of susceptibility to IPA infection. SNP-SNP interaction (epistasis) analysis revealed significant interactions models including SNPs in Dectin-1, Dectin-2, CCL2 and CCR2 genes, with synergistic genetic effects. Although these results need to be further validated in larger cohorts, they suggest that Dectin-1, DC-SIGN, Dectin-2, CCL2 and CCR2 genetic variants influence the risk of IPA infection and might be useful in developing a risk-adapted prophylaxis.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus fumigatus; Cell Adhesion Molecules; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Genotype; Humans; Lectins, C-Type; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Odds Ratio; Polymerase Chain Reaction; Polymorphism, Genetic; Polymorphism, Single Nucleotide; Receptors, Cell Surface; Risk

The serum galactomannan assay (GMA) has been widely used for the diagnosis of invasive aspergillosis (IA). GMA is mainly used in patients with haematological malignancies or in those who have undergone haematopoietic stem cell transplantation (HSCT). However, there are few data from non-haematological patients. We evaluated whether GMA is useful for the diagnosis of IA in non-haematological patients.. Patients who were subjected to serum GMA testing from January 2007 to December 2009 were evaluated retrospectively. Patients with haematological diseases or who underwent HSCT were excluded from our analysis. According to the criteria of the European Organization for Research and Treatment of Cancer/Mycoses Study Group revised in 2008, the patients were categorized as proven, probable, possible, or non-IA. Proven and probable cases were defined as IA in this study.. Out of 778 patients, 13 (1.6%) had proven (n =9) or probable (n =4) IA. The sensitivity of the GMA was 23.1% (95% confidence interval (CI) 6.1-54.0%) and the specificity was 76.1% (95% CI 72.9-79.0%). The positive predictive value was 1.6% (95% CI 0.4-5.0%) and the negative predictive value was 98.3% (95% CI 96.8-99.1%). The likelihood ratios of a positive and negative test were 0.96 (95% CI 0.35-2.62) and 1.01 (95% CI 0.75-1.36), respectively.. In this study, the sensitivity of the GMA for the diagnosis of IA was very low in non-haematological patients. Although the GMA test is considered useful for the diagnosis of IA in haematological patients, it had low diagnostic value for IA in non-haematological patients.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Female; Galactose; Humans; Immunoenzyme Techniques; Male; Mannans; Middle Aged; Predictive Value of Tests; Retrospective Studies; Sensitivity and Specificity

Galactomannan (GM) testing is extremely useful for diagnosing invasive aspergillosis in high-risk patients, but false-positive results have been reported in patients treated with piperacillin/tazobactam. The aims of this study are to test if the recent piperacillin/tazobactam (Tazocin™; Pfizer) preparation still contains GM, and if serum GM positivity in haematopoietic stem cell transplant (HSCT) recipients receiving piperacillin/tazobactam can be attributed to this treatment.. Serum samples obtained from 1 October 2009 to 31 October 2010 from HSCT recipients for GM testing were analysed. The difference in the rate of positive results (defined as GM ≥ 0.5) in patients receiving and not receiving piperacillin/tazobactam was evaluated. Piperacillin/tazobactam vials from randomly selected batches were tested.. Of 1606 samples drawn in the absence of piperacillin/tazobactam therapy, 25 (1.6%) tested positive for GM versus 10 of 394 samples (2.5%) drawn while on piperacillin/tazobactam (P = 0.18). The median GM result of samples drawn on piperacillin/tazobactam was slightly higher than that of samples drawn in the absence of piperacillin/tazobactam (0.141 versus 0.122; P < 0.001). All 90 piperacillin/tazobactam vials from 30 randomly selected batches tested negative for GM, with a median GM value of 0.057 (range: 0.011-0.320).. Although some residual GM might still be present in piperacillin/tazobactam, currently available brand piperacillin/tazobactam preparations seem no longer responsible for false-positive GM results.

Topics: Anti-Bacterial Agents; Aspergillosis; False Positive Reactions; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination

Cystic fibrosis patients are often colonized by Aspergillus species. Sera from 138 pediatric and adult cystic fibrosis patients were tested for the presence of galactomannan. All serum samples were negative for galactomannan and there was no difference among patients who were chronically, intermittently, and never colonized with Aspergillus.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Child; Cystic Fibrosis; Female; Forced Expiratory Volume; Galactose; Humans; Immunoenzyme Techniques; Lung; Male; Mannans; Middle Aged; Young Adult

Sera samples from commercial broiler chickens and turkeys diagnosed with respiratory and disseminated aspergillosis were tested for the presence of antigen and antibody to Aspergillus. Antigen detection consisted of testing for two cell-wall components, beta-glucan and galactomannan, which have been used extensively in human medicine. There were significantly higher levels of galactomannan in all broiler chicken submissions (100%) and antibody to Aspergillus in 6 out of 9 submissions (66.6%) vs. control birds. Beta-glucan analyses did not show any differences among levels in the broiler chicken groups. There were significantly higher levels of galactomannan antigen in 4 out of 7 submissions (57.1%) of aspergillosis in commercial turkeys, while only 2 out of 7 submissions (28.5%) had higher levels of antibody to Aspergillus vs. the control group. This study shows that diagnosis of respiratory and disseminated aspergillosis may be performed by detection of galactomannan antigenemia and antibodies in broiler chickens and to an extent in turkeys.

Topics: Animals; Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; California; Chickens; Enzyme-Linked Immunosorbent Assay; Galactose; Mannans; Poultry Diseases; Serologic Tests; Species Specificity; Turkeys

The presence of Aspergillus antigens in blood transfusion components from different manufacturers was analyzed. Galacomannans were found in transfused patients, pooled platelet concentrates, fresh frozen plasma, and packed red cells collected using Fresenius Kabi bags. Galacomannans were also found in blood collection anticoagulant and platelet additive solution from this manufacturer.

Topics: Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Cytomegalovirus Infections; False Positive Reactions; Female; Fungemia; Galactose; Humans; Mannans; Platelet Transfusion

Itraconazole is used for the prevention and treatment of infections caused by Aspergillus fumigatus. An understanding of the pharmacodynamics of itraconazole against wild-type and triazole-resistant strains provides a basis for innovative therapeutic strategies for treatment of infections. An in vitro model of the human alveolus was used to define the pharmacodynamics of itraconazole. Galactomannan was used as a biomarker. The effect of systemic and airway administration of itraconazole was assessed, as was a combination of itraconazole administered to the airway and systemically administered 5FC. Systemically administered itraconazole against the wild type induced a concentration-dependent decline in galactomannan in the alveolar and endothelial compartments. No exposure-response relationships were apparent for the L98H, M220T, or G138C mutant. The administration of itraconazole to the airway resulted in comparable exposure-response relationships to those observed with systemic therapy. This was achieved without detectable concentrations of drug within the endothelial compartment. The airway administration of itraconazole resulted in a definite but submaximal effect in the endothelial compartment against the L98H mutant. The administration of 5FC resulted in a concentration-dependent decline in galactomannan in both the alveolar and endothelial compartments. The combination of airway administration of itraconazole and systemically administered 5FC was additive. Systemic administration of itraconazole is ineffective against Cyp51 mutants. The airway administration of itraconazole is effective for the treatment of wild-type strains and appears to have some activity against the L98H mutants. Combination with other agents, such as 5FC, may enable the attainment of near-maximal antifungal activity.

Topics: Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Cells, Cultured; Drug Administration Routes; Drug Resistance, Fungal; Flucytosine; Galactose; Humans; Itraconazole; Lung Diseases, Fungal; Mannans; Microbial Sensitivity Tests; Pulmonary Alveoli; Triazoles

Invasive aspergillosis (IA) is the most common life-threatening infections after hematopoietic stem cell transplant (HSCT). The serum galactomannan (GM) is recognized as an indirect mycological criteria for an early diagnosis of IA. Starting January 2011, we implementing in Fundeni Clinical Institute, Bucharest, for the first time in Romania, the detection of GM antigen (Platelia Aspergillus EIA, Bio-Rad). In 2011, patients undergoing HSCT were screened with the galactomannan ELISA; we performed a retrospective chart review of 162 SCT patients who underwent galactomannan testing. Thirteen of the patients (8.02%) had at least one positive galactomannan ELISA, and four had multiple positive tests. When calculated in reference to a proved or probable diagnosis of aspergillosis, the galactomannan ELISA had a sensitivity of 0.857 and a specificity of 0.913. The positive predictive value was 0.46, and the negative predictive value was 0.993. The Platelia Aspergillus galactomannan antigenemia assay may assist physicians in making an early diagnosis of IA, in correlation with clinical and radiological criteria. The test has a high sensitivity and specificity and a very good negative predictive value. We found the screening of GM ELISA to be a highly specific diagnostic tool in detecting IA manifested in patients undergoing HSCT.

Topics: Adolescent; Adult; Aged; Aspergillosis; Child; Child, Preschool; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunoassay; Infant; Mannans; Middle Aged; Young Adult

Mortality associated with invasive aspergillosis (IA) remains high, partly because of delayed diagnosis. Detection of microbial exoantigens, released in serum and other body fluids during infection, may help timely diagnosis. In course of IA, Aspergillus galactomannan (GM), a well established polysaccharide biomarker, is released in body fluids including urine. Urine is an abundant, safely collected specimen, well-suited for point-of-care (POC) testing, which could play an increasing role in screening for early disease. Our main objective was to demonstrate GM antigenuria as a clinically relevant biological phenomenon in IA and establish proof-of-concept that it could be translated to POC diagnosis. Utilizing a novel IgM monoclonal antibody (MAb476) that recognizes GM-like antigens from Aspergillus and other molds, we demonstrated antigenuria in an experimental animal IA model (guinea pig), as well as in human patients. In addition, we investigated the chemical nature of the urinary excreted antigen in human samples, characterized antigen detection in urine by immunoassays, described a putative assay inhibitor in urine, and indicated means of alleviation of the inhibition. We also designed and used a lateral flow immunochromatographic assay to detect urinary excreted antigen in a limited number of IA patient urine samples. In this study, we establish that POC diagnosis of IA based on urinary GM detection is feasible. Prospective studies will be necessary to establish the performance characteristics of an optimized device and define its optimal clinical use.

Topics: Animals; Antibodies, Monoclonal; Antibody Specificity; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Cross Reactions; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Mice

Invasive fungal disease (IFD) is a leading cause of infection-related mortality among patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although the mortality of IFD has been decreased with empirical antifungal therapy in this population, overtreatment remains a problem, for the persistent or recurrent fever is nonspecific for an IFD. Hence, we explored retrospectively the value of incorporating serum galactomannan (GM) test and pulmonary high-resolution computed tomography (HRCT) as predictors for higher response rate of empirical treatment. In our study, all of 124 patients after allo-HSCT received empirical antifungal therapy when the persistent or recurrent fever developed after receiving broad-spectrum antibiotics. Meanwhile, the levels of serum GM were monitored twice weekly and pulmonary HRCT was performed serially. It showed that the response rate of empirical antifungal therapy was higher in patients with positive results of serum GM tests and/or chest CT scan than that in all patients (66.1% versus 44.6%, P = .008). Moreover, only 10 of 55 patients with both negative GM tests and pulmonary HRCT responded to empirical treatment, and 7 of these 10 patients did not take antifungal agents for prophylaxis. It suggested that these 2 diagnostic tools could not predict patients without adequate Candida prophylaxis well. Thus, we concluded that serum GM assays and pulmonary CT scan could find out patients who really need empirical antifungal therapy, which resulted in improving the efficiency of the treatment.

Topics: Adult; Antifungal Agents; Aspergillosis; Aspergillus; Child; Enzyme-Linked Immunosorbent Assay; Female; Fever; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Lung; Male; Mannans; Predictive Value of Tests; Tomography, X-Ray Computed; Transplantation, Homologous

The galactomannan assay to diagnose invasive aspergillosis is recommended and clinically utilized, yet the mechanism of galactomannan release from Aspergillus fumigatus is unknown. We used an A. fumigatus strain lacking calcineurin A (cnaA), already shown to be critically important for pathogenicity, to evaluate galactomannan kinetics. During the logarithmic growth phase when glucose was consumed, β-D-galactofuranoside (galf)-antigens were released in the culture. However, after glucose became limited, GM release further increased in the supernatants of the wild type strain while there was no further increase of GM release in the ΔcnaA strain. β-Galactofuranosidase activity was also decreased in the ΔcnaA mutant, and the amount of galf-antigen in the cell wall fraction of the ΔcnaA mutant was approximately ten-fold higher. This suggests the possibility that the antigen is unable to be released due to a cell wall abnormality. This and previous work suggest a dynamic calcineurin-dependent cell wall during periods of growth, with galactomannan release from the cell wall possibly calcineurin-dependent and reflected in the decreased GM release and greatly increased cell wall fraction of galf in the ΔcnaA mutant.

Topics: Aspergillosis; Aspergillus fumigatus; Calcineurin; Cell Wall; Culture Media; Galactose; Glucose; Glycoside Hydrolases; Humans; Mannans; Mutation

PCR assays designed for the diagnosis of invasive aspergillosis (IA) in high-risk patients have to detect minute amounts of target DNA to reach sufficient analytical sensitivity to be of clinical use. This prospective study assessed the use of a novel strategy for selective pathogen DNA enrichment for enhancing the performance of diagnostic PCR in a direct comparison with a highly sensitive in-house quantitative PCR (qPCR) assay and the galactomannan enzyme-linked immunosorbent assay (ELISA). Surprisingly, and in contrast to experience with other patient groups, the novel protocol for selective pathogen DNA enrichment did not enhance but instead significantly impaired sensitivity. This could be explained by the small amounts of host DNA in the specimens, which were derived mostly from severely neutropenic patients. In the qPCR assay, positive samples required an average of 43.5 amplification cycles (range, 39.2 to 50) for detection in the in-house PCR. Repetitive testing of selected samples showed test positivity to be variable, most likely due to the small amounts of target DNA. Despite this, the in-house protocol proved helpful in the diagnosis of IA, detecting 2 out of 3 patients with probable IA and 10 out of 19 patients with possible IA. Our results underline the necessity for diagnostic PCR protocols that help diagnose IA to be highly sensitive and show that selective pathogen DNA enrichment using affinity purification may not be useful in severely neutropenic patients.

Topics: Adult; Aged; Aspergillosis; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Mycology; Neutropenia; Polymerase Chain Reaction; Prospective Studies; Sensitivity and Specificity; Specimen Handling

β-D-(1,3)-glucan (BG) is a component of the cell walls of many fungal organisms. Our aims were to investigate the feasibility of the BG assay and its contribution to early diagnosis of different types of invasive fungal infections (IFI) commonly diagnosed in a tertiary care centre. The BG serum levels of 28 patients diagnosed with six IFI [13 probable invasive aspergillosis (IA), 2 proven IA, 2 zygomycosis, 3 fusariosis, 3 cryptococcosis, 3 candidaemia and 2 pneumocystosis] were retrospectively evaluated. The kinetic variations in BG serum levels from the 15 patients diagnosed with IA were compared with those of the galactomannan antigen (GM). In 5/15 cases of IA, BG was positive earlier than GM (time lapse from 4 to 30 days), in 8/15 cases, BG was positive at the same time as GM and, in 2/15 cases, BG was positive after GM. For the five other fungal diseases, BG was highly positive at the period of diagnosis except for the two cases of zygomycosis and one of the three cases of fusariosis. This study, which reflects the common activity of a tertiary care centre, confirms that BG detection could be of interest for IFI screening in patients with haematological malignancies.

Topics: Aspergillosis; beta-Glucans; France; Galactose; Hematologic Neoplasms; Humans; Immunocompromised Host; Mannans; Mycoses; Predictive Value of Tests; Proteoglycans; Retrospective Studies; Time Factors

Galactofuranose is a hexose that is exclusively found in microbes and in particular in certain pathogenic species. In the mold Aspergillus fumigatus, it is the characteristic constituent of the cell wall component galactomannan. Detection of this carbohydrate is currently a widespread method used for diagnosis of systemic A. fumigatus infections. In this study, we raised and characterized 2 monoclonal antibodies that specifically react with galactofuranose-containing glycostructures. We investigated the distribution of surface-accessible galactomannan on different A. fumigatus morphotypes. We provide evidence that the antibodies recognize distinct antigens and are suitable to detect A. fumigatus hyphae in immunohistology. A mutant that is impaired in synthesis of galactofuranose stimulated a normal cytokine response in murine macrophages, which argues against galactomannan being a relevant PAMP, at least in mice. Purified galactomannan-specific monoclonal IgM L10-1 failed to inhibit the hyphal growth under in vitro conditions, but L10-1 binding to hyphae led to an enhanced deposition of the complement protein C1q. However, administration of purified L10-1 antibodies prior to infection was not able to protect mice. In conclusion, we have found no evidence for galactomannan being a relevant A. fumigatus PAMP and describe 2 novel galactomannan antibodies that might be valuable tools for the diagnosis of A. fumigatus infections and further analysis of the biological significance of galactomannan.

Topics: Animals; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Cell Wall; Complement C1q; Electrophoresis, Polyacrylamide Gel; Galactose; Humans; Hybridomas; Hyphae; Immunoglobulin M; Interleukin-10; Macrophages; Male; Mannans; Mice; Mice, Inbred BALB C; Mutation

Myceliophthora thermophila is a thermophilic mould widely found in the environment but rarely responsible for human infections. We describe a case of invasive Myceliophthora thermophila infection mimicking invasive aspergillosis in a neutropenic patient with haematological malignancy. Cross-reactivity with Aspergillus galactomannan assay (GM) was demonstrated by repeated positive results and confirmed by cross-reaction between the fungal isolate and the GM assay. The patient was successfully treated with voriconazole. Potential GM cross-reactivity must be considered in future studies including patients categorized as having probable invasive aspergillosis using the GM as the only mycological criterion.

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Base Sequence; Cross Reactions; Diagnosis, Differential; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Molecular Sequence Data; Mycoses; Neutropenia; Pyrimidines; Sensitivity and Specificity; Sordariales; Spores, Fungal; Triazoles; Voriconazole

Topics: Adult; Aspergillosis; Aspergillus; Clinical Laboratory Techniques; Enteral Nutrition; False Positive Reactions; Female; Galactose; Gastrointestinal Diseases; Humans; Mannans; Mycology

The diagnosis of invasive aspergillosis remains very difficult, and there are limited treatment options for the disease. Pre-clinical models have been used to evaluate the diagnosis and treatment of Aspergillus infection and to assess the pathogenicity and virulence of the organism. Extensive efforts in Aspergillus research have significantly expanded the genomic information about this microorganism. The standardization of animal models of invasive aspergillosis can be used to enhance the evaluation of genomic information about the organism to improve the diagnosis and treatment of invasive aspergillosis. One approach to this process has been the award of a contract by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health to establish and standardize animal models of invasive aspergillosis for the development of new diagnostic technologies for both pulmonary and disseminated Aspergillus infection. This work utilizes molecular approaches for the genetic manipulation of Aspergillus strains that can be tested in animal-model systems to establish new diagnostic targets and tools. Studies have evaluated the performance characteristics of assays for cell-wall antigens of Aspergillus including galactomannan and beta-D-glucan, as well as for DNA targets in the organism, through PCR. New targets, such as proteomic and genomic approaches, and novel detection methods, such as point-of-care lateral-flow devices, have also been evaluated. The goal of this paper is to provide a framework for evaluating genomic targets in animal models to improve the diagnosis and treatment of invasive aspergillosis toward ultimately improving the outcomes for patients with this frequently fatal infection.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus; Biomarkers; Disease Models, Animal; DNA, Fungal; Galactose; Humans; Invasive Pulmonary Aspergillosis; Mannans; Predictive Value of Tests; Prognosis; Severity of Illness Index; Virulence Factors

A 66-year-old male with ischaemic cardiomyopathy and chronic lymphocytic leukemia developed signs of severe systemic inflammatory response syndrome. Serial blood cultures were negative and a SeptiFast test detected the presence of Aspergillus fumigatus DNA. Afterwards, detection of galactomannan and 1,3-β-D-glucan showed a positive result. Autopsy revealed the presence of branched fungal structures suggestive of Aspergillus.

Topics: Aged; Aspergillosis; Aspergillus fumigatus; Autopsy; beta-Glucans; Cardiomyopathies; DNA, Fungal; Endocarditis; Fatal Outcome; Galactose; Histocytochemistry; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Male; Mannans; Microscopy; Proteoglycans

Invasive aspergillosis is a life-threatening infectious complication in hematological patients undergoing immunosuppressive chemotherapy.. We report 29 cases of invasive aspergillosis diagnosed in the Sousse Farhat Hached hospital Hematology unit, Tunisia, between 2002 and 2010.. The most frequent disease (65.5%) was acute myeloid leukemia. All patients were severely neutropenic (<500/mm(3), mean duration=27 days). Pulmonary invasive aspergillosis was suggested in 28 (96.5%) cases. The most frequent respiratory signs were cough (64.3%), chest pain (53.6%), and hemoptysis (50%). The chest X-ray showed suggestive lesions in 60.7% of cases. CT scans revealed nodules with cavitation in 65% of cases, a halo sign in 20% of cases, and nodules in 15% of cases. Galactomannan antigenemia was positive in 88%, mycological examination positive in 51.6%, and seroconversion was noted in 35.7% of the cases. Invasive pulmonary aspergillosis was classified, according to EORTC/MSG criteria, as probable in 26 cases, possible in one case, and proven in one case. Aspergillus flavus was the dominant species in pulmonary invasive aspergillosis accounting for 73.7% of isolates. Extrapulmonary involvement was suggested in 39.3% of cases, the most frequent were sinusitis and brain abscess. Primary cutaneous aspergillosis was observed in one case. The overall mortality rate was 64.2%; the 12-week survival rate was 71.4%.. Our results are correlated to published data. A. flavus was the most frequent species in our region.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Antineoplastic Combined Chemotherapy Protocols; Aspergillosis; Aspergillus; Brain Abscess; Child; Child, Preschool; Dermatomycoses; Enzyme-Linked Immunosorbent Assay; Female; Fungemia; Galactose; Hematologic Neoplasms; Humans; Induction Chemotherapy; Invasive Pulmonary Aspergillosis; Male; Mannans; Middle Aged; Neuroaspergillosis; Neutropenia; Sinusitis; Survival Rate; Tomography, X-Ray Computed; Tunisia; Young Adult

A study was performed to investigate the air quality of a haematopoietic SCT ward, colonization of the upper airways with Aspergillus spp. and the role of galactomannan (GM) ELISA testing in serum in the diagnosis of invasive aspergillosis (IA). In 102 allo-SCT recipients, two cases of IA (one proven and one probable) were seen. Of 2071 serum samples, 12 were positive, two in a patient with proven IA and 10 in patients without IA. Of the 2059 negative samples, 22 were taken from the patient with probable IA. Of the 245 environmental samples, 20 (8.2%) were positive for filamentous fungi. Aspergillus fumigatus was seen in 14 samples. A total of 657 oral and nasal swabs were taken. Seven nasal samples and one oral sample were positive for Aspergillus species (A. fumigatus 4, A. niger 4) in four patients, one of whom had probable IA. In summary, most environmental samples were negative, colonization of the oral and nasal cavities was rare and IA was diagnosed in only 2% of patients. The GM ELISA test remained negative in one of two patients with IA and does not seem useful in a population of patients with a low incidence of IA.

Topics: Air Microbiology; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Enzyme-Linked Immunosorbent Assay; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Mouth; Nose; Patient Isolation

Aspergillus fumigatus is the major cause of invasive aspergillosis (IA), a disease associated with high rates of morbidity and mortality in patients undergoing treatment for haematological malignancies. This study investigated A. fumigatus growth in vitro and in a murine model of IA in order to provide insights into the dynamics of extracellular DNA and galactomannan (GM) release and their relevance to early diagnosis of IA. Following inoculation of whole blood with 20 A. fumigatus conidia ml(-1), DNA that corresponded to the inoculum could be detected by PCR but GM was not detected in plasma separated from the blood sample, indicating that the fungus did not grow in whole blood. The quantities of DNA detected by PCR, and GM, were proportional to the amount of fungal biomass present in vitro. Fungal DNA could be detected in the sera of mice experimentally infected with A. fumigatus with maximum detection in cyclophosphamide-treated mice.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Culture Media; DNA, Fungal; Galactose; Humans; Mannans; Mice; Mice, Inbred C57BL; Polymerase Chain Reaction

Invasive aspergillosis (IA) presents a diagnostic and therapeutic dilemma for the physicians who take care of the patients with severe underlying diseases and immunosuppression. This study aimed to evaluate the usefulness of serum galactomannan (GM) measurements in the routine practice and surveillance of IA along with possible caveats in diagnosis and treatment. Adult patients with high-risk haematological malignancies admitted to the Internal Medicine wards during the 2-year study period were followed up by daily visits for vital signs, existing or newly developing signs and symptoms, clinical and laboratory findings. Blood samples were analysed for GM levels by the ELISA method at the end of the study period. Data of 58 hospitalisation episodes in 45 patients were analysed. Proven IA was diagnosed in one patient, probable IA was diagnosed in four patients. The sensitivity was 60% and the specificity was 21% when the index cut-off for positivity was accepted as 0.5. The yield of GM testing may be influenced by many variables and each centre should evaluate the usefulness of this test in its own conditions.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Aspergillosis; Clinical Laboratory Techniques; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Neoplasms; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Population Surveillance; Sensitivity and Specificity; Young Adult

Cutaneous models have proven useful in studies of the pathogenesis and treatment of Gram-positive bacterial infections. Because cutaneous invasive aspergillosis (IA) occurs in the clinical setting, we sought to develop a nonlethal murine cutaneous model of IA. We induced cutaneous IA in cyclophosphamide-treated nude BALB/c mice by subcutaneous injection of Aspergillus fumigatus conidia. Skin lesion areas correlated well with tissue fungal burdens, allowing dynamic visual monitoring of cutaneous infections. The cutaneous model accurately reflected alterations in A. fumigatus pathogenicity resulting from deletions of recognized virulence genes (pabaA, sidA, and pksP). Moreover, analysis of the roles of conidial and mycelial catalases revealed that the former is required for the initiation of cutaneous aspergillosis, whereas the latter contributes to its propagation. Finally, posaconazole treatment reduced skin lesion areas relative to those of untreated and fluconazole-treated controls. This novel cutaneous model system should be applicable to comparative studies of the pathogenesis, treatment, and tissue specificity of IA.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Catalase; Cyclophosphamide; Dermatomycoses; Disease Models, Animal; Female; Galactose; Hyphae; Immunocompromised Host; Immunosuppressive Agents; Mannans; Mice; Mice, Inbred BALB C; Mice, Nude; Mutagenesis; Skin; Spores; Triazoles; Virulence; Virulence Factors

Prognostic features of serum galactomannan (GM) remain poorly defined in patients with GM-positive invasive aspergillosis (GPA). We identified 93 patients with proven or probable invasive aspergillosis (IA) and GM values of >or=0.50 from January 2005 to March 2009. We used Cox modeling of time to 6- and 12-week mortality for the GM level at the time of diagnosis (GM(0)), GM decay in the week following diagnosis in 72 patients with >or=2 GM values, other predictors of mortality, and antifungal use during the week following diagnosis. Six-week mortality was 55% in the whole cohort and 43% in patients with >or=2 GM determinations. The hazard ratio (HR) of GM(0) per unit increase and 1-week GM decay per unit decline per week were 1.25 (95% confidence interval [CI], 1.01 to 1.54; P = 0.04) and 0.78 (95% CI, 0.63 to 0.96; P = 0.02), respectively, adjusting for other predictors of IA mortality; these values remained stable after adjusting for antifungal use and were predictive of all-cause mortality at 12 weeks with similar adjusted HR values. We conclude that the combination of GM(0) and 1-week GM decay is predictive of all-cause mortality in patients with GPA, independent of other traditional risk factors for mortality and antifungal exposure, supporting GM decay as a potential surrogate endpoint for future antifungal therapeutic trials.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antifungal Agents; Antigens, Fungal; Aspergillosis; Female; Galactose; Humans; Immunoassay; Male; Mannans; Middle Aged; Prognosis; Serum; Time Factors; Treatment Outcome; Young Adult

Serum galactomannan (GM) antigen detection is not recommended for defining invasive aspergillosis (IA) in children undergoing aggressive chemotherapy or allogeneic haemopoietic stem cell transplantation (HSCT). The ability of the GM test to identify IA in children was retrospectively evaluated in a cohort of children. Test performance was evaluated on samples that were collected during 195 periods at risk of IA. Proven IA was diagnosed in seven periods, all with positive GM test results (true positives, 4%), and possible IA was diagnosed in 15 periods, all with negative GM test results (false negatives, 8%). The test result was positive with negative microbiological, histological and clinical features in three periods (false positives, 1%), and in 170 periods it was negative with negative microbiological, histological and clinical features (true negatives, 87%). The sensitivity was 0.32 and the specificity was 0.98; the positive predictive value was 0.70 and the negative predictive value was 0.92. The efficiency of the test was 0.91, the positive likelihood ratio was 18.3, and the negative likelihood ratio was 1.4. The probability of missing an IA because of a negative test result was 0.03. Test performance proved to be better during at-risk periods following chemotherapy than in periods following allogeneic HSCT. The GM assay is useful for identifying periods of IA in children undergoing aggressive chemotherapy or allogeneic HSCT.

Topics: Adolescent; Aspergillosis; Child; Child, Preschool; Galactose; Humans; Immunoassay; Immunocompromised Host; Infant; Mannans; Mycology; Neoplasms; Predictive Value of Tests; Retrospective Studies; Sensitivity and Specificity; Stem Cell Transplantation; Young Adult

Our objective was to identify false-positive serum and bronchoalveolar lavage (BAL) fluid galactomannan (GM) tests caused by various antibiotics commonly used in general practice. Serum and BAL samples from patients who did not have the diagnostic criteria of invasive aspergillosis and received different antibiotics were prospectively analyzed for GM. Serum and BAL samples were also collected from patients who did not receive antibiotics. At the cut-off index of >or=0.5, false-positive serum results were found in patients who received amoxicillin-clavulanate, piperacillin-tazobactam, cefepime, and cefoperazone-sulbactam (26.7%, 58.3%, 14.3%, and 66.7%, respectively). Fungal colonization in BAL samples had a higher BAL GM than those without fungal colonization. In 71 patients who had a negative BAL culture for fungi, at the cut-off value of >or=1.0, false-positive BAL fluid results were found in patients who received amoxicillin-clavulanate (27.3%), piperacillin-tazobactam (50%), cefepime (16.7%), carbapenem (45.5%), and ceftriaxone (45.5%). False-positive serum and BAL GM assays were also detected in patients who did not receive any antibiotics. In summary, this study demonstrates the false-positive GM levels in serum and BAL caused by beta-lactam antibiotics that are commonly used in general practice. Physicians should be aware of this possible interference.

Topics: Adult; Aged; Anti-Bacterial Agents; Antibiotic Prophylaxis; Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Chi-Square Distribution; Disease; False Positive Reactions; Female; Galactose; Humans; Male; Mannans; Middle Aged; Predictive Value of Tests; Prospective Studies; Statistics, Nonparametric

The aim of this study was to investigate the interaction between intravenous piperacillin/tazobactam treatment and Aspergillus galactomannan antigen (GM) and 1,3-beta-D: -glucan (BDG) test results in patients without known risk factors for invasive fungal infections (IFI).. Patients without known risk factors for IFI and who were to receive piperacillin/tazobactam monotherapy were considered eligible for the study. Serum samples were obtained both before and after antibiotic infusion on the first, third, seventh and tenth days of a piperacillin/tazobactam treatment course and 4 days after the last dose. GM was determined by Platelia Aspergillus ELISA (Bio-Rad Laboratories) and BDG was assayed using the Fungitell kit (Associates of Cape Cod, East Falmouth, MA) according to manufacturers' specifications.. A total of 135 serum samples were collected from 15 patients. When a cut-off level of >or=0.7 was used for GM positivity, there were no false positive results. When a cut-off level of >or=0.5 was used, six serum samples were positive. There were no statistically significant differences between the median GM indices or median BDG levels of the various sampling times. However, 24 of 135 serum samples were positive for BDG for a threshold of 80 pg/mL. After ruling out fungal infections and all known potential causes of false BDG positivity, environmental contamination remained a possible cause of BDG reactivity.. No significant interaction was observed between piperacillin/tazobactam administration and Aspergillus GM and BDG assays. Positive results for these tests should be evaluated cautiously in patients at high risk for IFI receiving piperacillin/tazobactam.

Topics: Adult; Aged; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; False Positive Reactions; Female; Galactose; Humans; Infusions, Intravenous; Male; Mannans; Microbiological Techniques; Middle Aged; Mycology; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Risk Factors

To evaluate the role of real-time polymerase chain reaction (real-time PCR) assay combined with serum galactomannan (GM) detection in the diagnosis of invasive aspergillosis (IA) with hematological malignancies or hematopoietic stem cell transplantation.. Eighty patients were enrolled. PCR and GM were performed simultaneously on 205 serum samples. The agreement between these two assays and the diagnostic value of the combination were analyzed.. There were 5 proven IA, 20 probable IA, 34 possible IA and 21 non-IA. Agreement between real-time PCR and GM was 77.5%. When both positive, the positive predictive value could reach 100%; when both negative, the negative predictive value could reach 90.9%.. PCR agrees well with GM. When these two assays are combined, invasive aspergillosis can be diagnosed more accurately.

Topics: Adolescent; Adult; Aged; Aspergillosis; Child; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Serum; Young Adult

PCR screening for circulating DNA, especially when combined with antigen testing, has shown promise for the definitive diagnosis of invasive aspergillosis. False positives for Aspergillus real-time PCR assays have been described in several reports, but no sources of fungal DNA contamination could be clearly identified. We report a false-positive case for both galactomannan (GM) antigenemia and Aspergillus PCR due to nutritional supplement intake in a bone marrow transplant recipient with digestive graft-versus-host disease. Our case report also suggests that fungal DNA can pass into the serum from the intestinal tract in the same way as fungal GM. Clinicians should be aware of this possibility, so that the administration of costly, unnecessary antifungal treatments with potential adverse side-effects can be avoided.

Topics: Adult; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Dietary Supplements; DNA, Fungal; False Positive Reactions; Galactose; Graft vs Host Disease; Humans; Immunocompromised Host; Male; Mannans; Reverse Transcriptase Polymerase Chain Reaction

Invasive aspergillosis (IA) remains one of the most significant causes of morbidity and mortality in patients with hematological malignancies undergoing chemotherapy and hematopoietic stem cell transplantation (HSCT), mainly due to the difficulty in its early diagnosis. Monitoring of galactomannan (GM) antigen, an exoantigen of Aspergillus, in the blood by sandwich ELISA is a useful and noninvasive method for early diagnosis of IA. The GM test has a sensitivity of 67-100% with a specificity of 81-99% in neutropenic patients and allogeneic transplant recipients [1-3]. Although it has been widely used as a diagnostic criterion for IA [4,5], one of the major limitations of this assay is false-positivity, particularly in pediatric patients [1], patients with graft-versus-host disease (GVHD) [6,7], and those taking dietary GM [8,9] or fungus-derived antibiotics, such as piperacillin-tazobactam (PIPC/TAZ) [10-12].

Topics: Amoxicillin; Antibiotic Prophylaxis; Antigens, Fungal; Artifacts; Aspergillosis; Aspergillus; False Positive Reactions; Galactose; Hematologic Diseases; Hematologic Neoplasms; Humans; Immunoglobulin G; Mannans; Multiple Myeloma; Myeloma Proteins; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Retrospective Studies; Sensitivity and Specificity

Topics: Acute Disease; Aspergillosis; Galactose; Humans; Male; Mannans; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Tomography, X-Ray Computed; Young Adult

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Glucans; Biomarkers; Colorimetry; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Nephelometry and Turbidimetry; Polymerase Chain Reaction; Proteoglycans

The European Organization for Research and Treatment of Cancer (EORTC) and the Mycosis Study Group (MSG) definition of invasive aspergillosis used in clinical trials lacks sensitivity. We hypothesize that giving lower weight to the prespecified radiologic findings in patients with a positive serum galactomannan index test result will improve the definition's diagnostic sensitivity.. The medical records of 121 patients with 125 cases of invasive aspergillosis treated at a referral cancer institute from January 2003 through December 2009 were reviewed. Aspergillosis was diagnosed as EORTC-MSG proven or probable (controls, 83) or probable invasive aspergillosis without prespecified radiologic criteria (cases, 42). The latter differed from the former by the inclusion of patients whose pulmonary infiltrates, although well described in invasive aspergillosis, do not fulfill EORTC-MSG invasive aspergillosis requirements. The host, clinical, and mycologic characteristics and survival of cases and controls served as end points.. A total of 114 (91%) of 125 patients had multiple myeloma. Patients had a median age was 65 years (range, 26-81 years), and 74 were male. All had received antineoplastic therapy, including stem cell transplantation (58 [46%]). Aspergillosis involved lungs (88 patients), sinuses (9 patients), or both (28 patients). Except for higher median baseline platelet count and shorter duration of neutropenia among cases, there were no statistically significant differences between groups on all predefined end points, including 4-, 6-, and 12-week survival. Eleven of 26 cases were reclassified as controls on the basis of subsequent imaging.. Except for less well-circumscribed consolidations, the host, clinical, radiologic, and mycologic characteristics and outcome of patients with probable invasive aspergillosis but without prespecified radiologic criteria are similar to those with EORTC-MSG invasive aspergillosis. Enrolling such patients in clinical trials of novel therapies will increase the pool of eligible study participants and improve trial speed and efficiency.

Topics: Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; Female; Galactose; Humans; Lung; Male; Mannans; Middle Aged; Paranasal Sinuses; Radiography; Retrospective Studies; Survival Analysis; Treatment Outcome

Topics: Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Lung; Mannans; Radiography, Thoracic; Sensitivity and Specificity; Tomography, X-Ray Computed

Invasive aspergillosis (IA) is a serious opportunistic infection in immunocompromised patients. Transplant recipients and patients with cancer represent the highest risk group. The antifungal treatment involves prolonged hospitalization and high economic resources.. to estimate costs represented by IA as an intercurrent complication of oncologic treatment.. Retrospective case-control study. Estimation of the cost of treatment in pediatric oncologic patients with IA in the Hospital Luis Calvo Mackenna during the years 2007-2008 was done. A control for each case of IA paired by sex, age, number of diagnosis and clinical department was selected.. There were 13 patients during the observation period. The attributable cost of treatment of aspergillosis was US $23,600 and the cost for each indicator was: hospital days US $16,500; antifungal therapy US $7,000; and serum galactomannan US $100.. In this study, the cost of treating IA is mainly due to hospitalization and antifungal medications. Three patients acquired IA in spite of staying in a protected environment.

Topics: Adolescent; Antifungal Agents; Antigens, Fungal; Aspergillosis; Case-Control Studies; Child; Chile; Cross Infection; Female; Galactose; Health Care Costs; Humans; Immunocompromised Host; Male; Mannans; Neoplasms; Opportunistic Infections; Retrospective Studies

In order to improve invasive pulmonary aspergillosis (IPA) diagnosis, a real-time polymerase chain reaction (PCR) assay detecting Aspergillus spp. was developed. Its detection limit reached 2-20 conidia. The retrospective evaluation on 64 bronchoalveolar lavage (BAL) fluids from 57 patients at risk for IPA, including 20 probable and five proven IPA patients, revealed a 88% or 38% sensitivity in direct examination (DE)/culture-positive or culture-negative BAL, respectively, whereas galactomannan (GM) sensitivity reached 88% or 58%, respectively. Influence on the Aspergillus-PCR yield of BAL fluid volume, cellular count and DNA content (evaluated by human beta-globin quantification) was assessed. Significantly higher beta-globin levels were detected in Aspergillus PCR-positive (median 5,112 pg/microl) than negative (median 1,332 pg/microl) BAL fluids, suggesting that the beta-globin level could reflect BAL yields and DNA extraction. Using beta-globin for the interpretation of fungal PCR could improve the negative predictive value of this test.

Topics: Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; DNA, Fungal; Female; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Predictive Value of Tests; Sensitivity and Specificity

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Plateliat test is unreliable for the diagnosis of canine SNA.

Topics: Animals; Antibodies, Fungal; Aspergillosis; Aspergillus; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Immunoglobulin G; Male; Mannans; Nose Diseases; Sinusitis

A study was designed to assess the reliability of the serial detection of Aspergillus sp. DNA to diagnose invasive aspergillosis (IA) in patients with febrile neutropenia. Two blood and two serum samples were taken weekly from 83 patients. A total of 2,244 samples were analyzed by real-time quantitative PCR. Twelve (14.4%) patients were diagnosed with IA. Taking two consecutive positive results as the diagnostic criterion, PCR detected 11 cases, with 4 false positives, giving sensitivity, specificity, positive, and negative predictive values of 91.6%, 94.4%, 73.3%, and 98.5%, respectively. On analyzing in conjunction with high-resolution chest tomography (HRCT) and galactomannan (GM) testing, the combination of serial PCR and GM detected 100% of aspergillosis cases, with a positive predictive value of 75.1%. This diagnostic strategy presented, according to CART analysis, a receiver-operator curve with an area under the curve of 0.97 (95% confidence interval, 0.895 to 1.032; P < 0.01), with a relative risk of IA 6.92 times higher than the control population and with predictive success of 95.2%. As regards early diagnosis, the serial detection of Aspergillus DNA took on average 21 days less than HRCT and 68 days less than GM. The serial detection of Aspergillus DNA using real-time quantitative PCR has great diagnostic applicability, which increases when combined with GM quantification.

Topics: Aspergillosis; Aspergillus; Blood; DNA, Fungal; Early Diagnosis; Female; Fever; Galactose; Humans; Male; Mannans; Middle Aged; Neutropenia; Polymerase Chain Reaction; Predictive Value of Tests; Radiography, Thoracic; ROC Curve; Sensitivity and Specificity; Serum; Time Factors

A noninvasive, objective, reproducible, and quantitative Aspergillus-specific surrogate marker is needed for a more accurate assessment of the outcome of invasive aspergillosis (IA) in patients with a hematologic disorder. The quantitative serum galactomannan index (GMI) assay seems to fulfill the requirements of surrogacy for outcome evaluation.. Kappa statistics were used to determine the strength of correlation between GMI outcome and clinical outcome (survival or death), autopsy data, and response outcome of IA in 70 adults with prolonged neutropenia. All patients underwent serial GMI monitoring until discharge or death.. The overall correlation between GMI and clinical outcome was good at 6 weeks (kappa=0.5882; 95% confidence interval [95% CI], 0.4023-0.7741) and was excellent at 12 weeks (kappa=0.8857; 95% CI, 0.7766-0.9948). Concordance with autopsy findings was perfect (kappa=1). At 6 weeks, the correlation between GMI and response outcome (favorable or unfavorable) was excellent (kappa=0.7523; 95% CI, 0.5803-0.9243). Survival was significantly better in patients who became GMI-negative (P<.0001).. In neutropenic patients with seropositive IA, serum galactomannan index outcome strongly correlates with survival, autopsy findings, and response outcome. This finding may have implications for patient management and for clinical trial design.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Autopsy; Biomarkers; Female; Galactose; Humans; Male; Mannans; Middle Aged; Neutropenia; Survival Analysis; Time Factors; Treatment Outcome

Topics: Adolescent; Aspergillosis; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Neoplasms; Predictive Value of Tests; Transplantation, Homologous

The detection of circulating galactomannan (GM) in serum samples is an important step in the diagnosis of invasive aspergillosis (IA). The assay has been mainly explored in neutropenic patients, and is now used to monitor patients at high risk for IA. However, the performance of the assay varies greatly among studies. The objective of this study was to explore the impact of the neutrophil count on the GM serum index at the time of IA diagnosis. Ninety-nine episodes of proven or probable, microbiologically documented IA in 91 patients with haematological malignancies were studied retrospectively. Three groups were identified: groups 1-3, with <100 polymorphonuclear neutrophils (PMN)/mm(3) (n = 18), between 100 and 500 PMN/mm(3) (n = 21), or >500 PMN/mm(3) (n = 60), respectively. The mean GM index was significantly higher in group 1 than in the other groups (p <0.05). This finding did not change after stratifying the analysis with regard to the use of antibiotics likely to give false-positive GM results or with regard to treatment effective against fungi before the diagnosis of IA. This finding could be considered in the routine use of the GM antigenaemia test in non-neutropenic patients; a negative result or a low GM index should not eliminate the diagnosis of IA. This limitation calls for other microbiological tests, including analysis of bronchoalveolar lavage fluid, to establish a definitive diagnosis of IA.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anti-Bacterial Agents; Antifungal Agents; Antigens, Fungal; Aspergillosis; Female; Galactose; Hematologic Neoplasms; Humans; Immunosuppressive Agents; Leukocyte Count; Male; Mannans; Middle Aged; Neutropenia; Neutrophils; Statistics, Nonparametric; Stem Cell Transplantation

Aspergillus galactomannan (GM) antigenemia is an early marker of invasive aspergillosis (IA), but may yield false-positive results. A prospective study, testing GM periodically in serum samples of liver transplant recipients, was performed.. An index more than or equal to 0.5 were considered positive. Positive GM in samples from patients without any other criteria of proven or probable IA was considered as false-positive. The test was performed weekly during the first month after transplantation.. Three patients developed IA. In total, 414 serum samples from 85 liver transplant recipients were analyzed. Mean number of samples per patient (out of those who could be assessed) was 4.8. The number of false-positive GM samples was 40 (9.6%), corresponding to 28 patients. The frequency of false-positive results in samples obtained during the first week posttransplantation was 36% (27 of 75), significantly higher than the number of false-positive samples obtained after the first week (3.8%; 13 of 339; P<0.001). Multivariate analysis showed that antibacterial prophylaxis with ampicillin was the only independent factor associated with a false-positive result. Different vials of beta-lactam antibiotics were tested for GM. We obtained a positive GM value (>0.5) in four of the six vials of ampicillin, in three of the six vials of piperacillin-tazobactam, in none of the six vials of cefotaxime, and in none of the six controls.. The present study suggests that the administration of ampicillin as antibacterial prophylaxis during the first days after transplantation could be a possible cause of false-positive GM results.

Topics: Ampicillin; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Drug Contamination; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Galactose; Humans; Liver Transplantation; Mannans; Predictive Value of Tests; Prospective Studies; Time Factors

Systemic histoplasmosis is uncommonly reported in patients who have undergone bone marrow or solid organ transplantation. Diagnosis of systemic histoplasmosis in recipients of transplants may be hampered by lack of consideration of this infection in the differential diagnosis and may be confounded by conflicting information from other testing performed to evaluate for opportunistic infections in this population. We report successful treatment of a case of disseminated histoplasmosis in a patient with Hodgkin's lymphoma who had undergone autologous stem cell transplantation. The diagnosis was delayed by the finding of a positive serum galactomannan assay.

Topics: Adult; Antifungal Agents; Aspergillosis; Diagnostic Errors; False Positive Reactions; Galactose; Histoplasma; Histoplasmosis; Hodgkin Disease; Humans; Male; Mannans; Stem Cell Transplantation; Transplantation, Autologous; Treatment Outcome

Previous studies support the possible application of galactomannan, a major antigen of Aspergillus sp., to aspergillosis diagnosis in avian and other animal species. An assay is commercially available for use with human serum and bronchoalveolar lavage fluid samples. In the current study, galactomannan results from plasma samples were compared between birds with histologically confirmed aspergillosis and those that were clinically normal presumptively non-Aspergillus infected birds per submitting practitioners' responses to a questionnaire. It was observed that infected birds demonstrated a 2.6-fold increase in galactomannan over birds without evidence of aspergillosis. With the use of a galactomannan index of 0.5 as a cutoff, the sensitivity and specificity of the test were found to be 67% and 73%, respectively. In addition, plasma samples were analyzed for abnormalities in protein electrophoretic patterns. Infected birds had a higher incidence of increased beta and/or gamma globulin concentrations. Test sensitivity and specificity were 73% and 70%, respectively. If the 2 tests were used as a panel, then the sensitivity was 89% and specificity was 48%. These data indicate that both galactomannan and protein electrophoresis may be valuable tools in the diagnosis of avian aspergillosis.

Topics: Animals; Animals, Domestic; Aspergillosis; Aspergillus; Bird Diseases; Birds; Diagnosis, Differential; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Galactose; Mannans; Sensitivity and Specificity; Surveys and Questionnaires; Veterinary Medicine

Topics: Aged, 80 and over; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Incidence; Japan; Male; Mannans; Pneumonia, Aspiration; Retrospective Studies

The objective of this study was to explore the useful value of circulating galactomannan (GM) for early diagnosis of invasive aspergillosis. All 141 patients were classified as 103 patients of clinical and possible diagnosis, and 38 non-Aspergillus patients. 209 serum samples for the detection of GM by Platelia Aspergillus were collected before anti-fungal vaccine therapy. ELISA method was used in detection of GM. The results showed that (1) the sensitivity of 87.5%, specificity of 81.6%, positive prediction of 66.7% and negative prediction of 93.9% were determined by using cut-off value. According to the result of ELISA, the clinical diagnosed patients was up to 48, while the possible diagnosed patients were 55. (2) Among 62 patients with consecutive examinations of serum samples, 50 patients were successfully diagnosed and treated, while 12 patients died. A progressive reduction of GM level was found in survivors, however, the patients of poor prognosis showed higher antigen titres. It is concluded that GM test has more significance for earlier diagnosis of aspergillosis, the concentration of GM is related to prognosis of disease.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Diseases; Humans; Male; Mannans; Middle Aged; Predictive Value of Tests; Sensitivity and Specificity; Young Adult

In psittacine birds, the antemortem diagnosis of aspergillosis is usually based on the clinical signalment combined with the results of diagnostic tests such as radiography, routine hematologic and biochemical analysis, and biopsy. For several years, plasma protein electrophoresis has been used as an ancillary diagnostic technique in forming a diagnosis and treatment plan in avian species. More recently, a commercially available assay to measure galactomannan, an Aspergillus species antigen, has been described for clinical use in humans, cattle, horses, dogs, and gyr falcons. This report describes several confirmed cases of aspergillosis, with accompanying clinical data, including plasma protein electrophoresis and galactomannan assay results, in addition to results of traditional evaluations by hematology, radiography, and biopsy. In clinical cases in psittacine birds, the galactomannan assay appears useful for detecting circulating Aspergillus antibody.

Topics: Animals; Aspergillosis; Bird Diseases; Blood Protein Electrophoresis; Female; Galactose; Male; Mannans; Psittaciformes

Invasive aspergillosis is a major cause of morbidity and mortality in immunocompromised patients receiving intensive care. The double-sandwich ELISA for galactomannan is reported to have a high sensitivity (96.5%) for the detection of invasive aspergillosis when a cut-off value of 0.8 ng/ml is used. However, we have experienced a case of lethal disseminated aspergillosis in a patient that presented with a negative galactomannan (GM) test and persistent elevation of beta-D glucan (BG) levels. A 63-year-old female was admitted to our Intensive Care Unit (ICU) in acute respiratory failure and elevated BG. She had been receiving medication for Good-pasture syndrome based on anti-glomerular basement membrane antibodies and myeloperoxidase-antineutrophil cytoplasmic antibodies for 9 months and was receiving long-term prednisolone therapy (20 mg/day). On admission, her trachea was immediately intubated, and a PCR analysis of the bronchoalveolar lavage sample revealed Pneumocystis jiroveci. Trimethoprimsulfamethoxazole therapy was started for Pneumocystis pneumonia. The levels of BG remained elevated (> 100 pg/ml) during the treatment period despite the clinical resolution of Pneumocystis pneumonia, raising concerns of another complicated invasive fungal disease; consequently, fosfluconazole was administered empirically. The serum BG levels, however, did not decrease. Blood cultures did not detect a fungal infection. Serum GM levels measured by a double-sandwich ELISA on the 6th, 11th, and 24th days in the ICU were negative (< 0.2 ng/ml). The patient ultimately died of multiple organ failure on the 45th ICU day. Postmortem examination revealed a disseminated fungal infection with aggressive vascular invasion of the lungs, heart, and brain. In situ hybridization with a 568-bp probe of the alkaline proteinase sequence of Aspergillus fumigatus showed specific positive staining within the fungus present in the infected lung tissue, revealing that this patient may have had a systemic infection by A. fumigatus or A. flavus. This is a case of serum GM-negative disseminated aspergillosis pathologically proven by autopsy. Persistent elevated BG levels (> 100 pg/ml) refractory to trimethoprim-sulfamethoxazole and fosfluconazole may suggest possible Aspergillus infection and should prompt the initiation of empiric anti-aspergillosis therapies in patients at risk for fungal infection.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; beta-Glucans; Brain; Fatal Outcome; Female; Galactose; Heart; Humans; Immunosuppressive Agents; Intensive Care Units; Lung; Mannans; Middle Aged; Pneumocystis carinii; Pneumonia, Pneumocystis; Prednisolone; Trimethoprim, Sulfamethoxazole Drug Combination

The diagnosis of invasive aspergillus remains a challenge in the care of high-risk patients. Outcomes are improved when invasive aspergillus is diagnosed early, prompting the initiation of appropriate antifungal therapy. We evaluated the utility of prospective monitoring for invasive aspergillosis (IA) using biomarkers such as serum galactomannan (GM) and/or blood polymerase chain reaction (PCR) in high-risk pediatric patients.. Patients with high-risk leukemia (HRL) or allogenic hematopoietic cell transplant (HCT) recipients were prospectively monitored twice weekly for IA using GM and PCR for Aspergillus species.. Sixty-eight patients had collected >or=2 specimens. The 1086 specimens were collected; 627 from HRL (58%) and 459 (42%) from HCT recipients. Median specimens/patient was 11.0 (2 to 58), and median follow-up/patient was 98.5 days (14 to 437). Fifty-six percent of samples were obtained from patients receiving mold-active agents; 32% HRL and 89% HCT. There were no proven, 3 probable, and 20 possible episodes of IA. Thirteen specimens (1.2%) from 4 patients (5%) were GM+. None were positive by PCR.. The prospective use of GM and PCR in this high-risk pediatric population did not identify cases of proven IA. A high false positive rate was not detected. It is speculated that changes in clinical practice, such as early use of empiric and/or prophylactic mold-active agent and frequent imaging studies have impacted the epidemiology of IA. In a population with low incidence of IA, the use of these assays as a screening device on blood may not further enhance current outcomes.

Topics: Adolescent; Adult; Aspergillosis; Aspergillus; Child; Child, Preschool; DNA, Fungal; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Feasibility Studies; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Infant; Male; Mannans; Polymerase Chain Reaction; Prospective Studies; Risk Factors; RNA, Ribosomal, 28S; Transplantation, Homologous; Young Adult

To explore the value of circulating galactomannan (GM) screening for early diagnosis and treatment monitoring of invasive aspergillosis (IA).. Serum samples from 141 IA patients for the detection of GM by Platelia Aspergillus (Bia-Rad) were collected before and after systematic anti-fungal therapy.. (1) An increase in the clinical diagnosis rate of IA was obtained on the result of GM detection. The GM positivity appeared (10+/-4.1) (8-15) d before positive sputum culture, while (12.6+/-5.7) (6-22) d before the CT positive image. (2) Among the 62 patients with consecutive serum samples, 50 were success in treatment and 12 died. A progressive decrease of GM level was found in the former group, while the rising antigen titres were found in the latter.. Compared with other diagnostic test, GM test has an obvious advantage of higher positivity and earlier result. The anti-fungal effectiveness can be estimated by dynamic detection of serum GM.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Young Adult

To analyse the predominant radiological pattern of pulmonary lesions in adult hematologic patients at risk for invasive aspergillosis (IA) together with the results of serial serum Aspergillus galactomannan antigen testing (GM).. In a prospective study for patients at high risk of aspergillus pulmonary infection, serum GM were performed 2-3 times per week during the periods of high risk for IA and high-resolution CT (HRCT) was performed in case of abnormal chest X-ray (CXR) and/or persistent fever after 5 days of antibiotic treatment. Changes on HRCT scan were classified as airway IA and angioinvasive IA. IA was classified as proven or probable in accordance with the definitions stated by the European Organization for Research and Treatment of Cancer/Mycosis Study Group (EORTC-MS). Positive GM testing was not considered as microbiological criterion.. 38 hematological patients were diagnosed of probable (n=28) or proven (n=10) IA. 55% patients had a neutrophil count less than 500 mm(-3) (n=21), and 37% patients > or =2 risk factors for IA. All probable IA were diagnosed by bronchoalveolar lavage (BAL). Proven IA was reached by positive histopathologic and culture results of samples obtained by autopsy (n=4), percutaneous (n=3) or transbronchial biopsy (n=3). 18 patients had airway IA, and 60% had a GM level > or =1.5. 20 patients were diagnosed of angioinvasive IA from which 80% had a GM level > or =1.5.. Serum GM levels may be lower in patients with airway IA than in those with an angioinvasive form. HRCT and serum GM are complementary tests in the diagnosis of IA.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; Female; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Reproducibility of Results; Risk Assessment; Sensitivity and Specificity; Statistics as Topic; Tomography, X-Ray Computed; Young Adult

Invasive mycoses represent a rare but severe complication following hemopoietic SCT (HSCT) in children. Their incidence is related to the type of donor, being higher after allogeneic transplant, especially from alternative donors. Moreover, the incidence of invasive mycoses varies in the different post transplant phases. Neutropenia, lymphopenia, GvHD, high-dose steroids or other immunosuppressive drugs represent well-known risk factors. The clinical features of invasive mycoses after HSCT in children are similar to those observed in adults, and the diagnostic tools, including Aspergillus galactomannan antigen detection, are feasible also in pediatrics. Mortality due to invasive mycoses after HSCT in children is high.

Topics: Aspergillosis; Child; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Mycoses; Risk Factors

The filamentous fungus Aspergillus fumigatus is responsible for a lethal disease called invasive aspergillosis that affects immunocompromised patients. This disease, like other human fungal diseases, is generally treated by compounds targeting the primary fungal cell membrane sterol. Recently, glucan synthesis inhibitors were added to the limited antifungal arsenal and encouraged the search for novel targets in cell wall biosynthesis. Although galactomannan is a major component of the A. fumigatus cell wall and extracellular matrix, the biosynthesis and role of galactomannan are currently unknown. By a targeted gene deletion approach, we demonstrate that UDP-galactopyranose mutase, a key enzyme of galactofuranose metabolism, controls the biosynthesis of galactomannan and galactofuranose containing glycoconjugates. The glfA deletion mutant generated in this study is devoid of galactofuranose and displays attenuated virulence in a low-dose mouse model of invasive aspergillosis that likely reflects the impaired growth of the mutant at mammalian body temperature. Furthermore, the absence of galactofuranose results in a thinner cell wall that correlates with an increased susceptibility to several antifungal agents. The UDP-galactopyranose mutase thus appears to be an appealing adjunct therapeutic target in combination with other drugs against A. fumigatus. Its absence from mammalian cells indeed offers a considerable advantage to achieve therapeutic selectivity.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Body Temperature; Cell Proliferation; Cell Wall; Disease Models, Animal; Drug Resistance, Fungal; Female; Furans; Galactose; Gene Expression Regulation, Fungal; Immunocompromised Host; Intramolecular Transferases; Mannans; Mice; Mice, Inbred BALB C; Opportunistic Infections; Virulence

Topics: Aspergillosis; False Positive Reactions; Galactose; Humans; Mannans

Invasive aspergillosis (IA) is a life-threatening complication of liver transplantation. Detection of circulating galactomannan (GM) in serum samples is a method to improve the microbiological diagnosis in patients at risk for IA. However, the assay is hampered by false-positive results. The search for circulating Aspergillus DNA in the first GM-positive sample could improve the specificity of the test. Among 484 liver transplant recipients followed in a single center over 4 years, 25 patients had at least 1 GM-positive serum sample. The threshold of GM positivity was a ratio >or=1. These 25 patients were classified by the clinicians as probable IA (n=11), possible IA (n=2), and no IA (n=12) using the EORTC/MSG criteria with blinding to the polymerase chain reaction (PCR) results. After 1 mL aliquots of the first GM-positive serum sample were thawed, 2 independent DNA extractions were performed using the MagNA Pure Compact apparatus. Real-time amplification targeted at Aspergillus fumigatus mitochondrial DNA was performed on 10 microL of the final eluate in duplicate in the 2 independent DNA extractions using a LightCycler instrument. A sample was considered positive when the crossing point was

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; DNA Primers; DNA, Fungal; Female; Galactose; Humans; Liver Transplantation; Male; Mannans; Middle Aged; Opportunistic Infections; Polymerase Chain Reaction; Retrospective Studies; Sensitivity and Specificity; Young Adult

False-positive results of the galactomannan (GM) ELISA caused by concurrent administration of piperacillin/tazobactam have been reported in patients with febrile neutropenia.. This prospective study investigated different sampling times in 30 patients receiving piperacillin/tazobactam for febrile neutropenia.. Prior to the first piperacillin/tazobactam infusion, a median GM index of 0.2 [interquartile range (IQR) 0.1-0.3] was noted; in two patients (7%) the index was 0.5. Immediately after piperacillin/tazobactam infusion, the median index increased to 0.3 (IQR 0.2-0.4, P = 0.002) leading to 21% (7/30) false-positive results, if > or = 0.5 is assumed as the cut-off level. GM indices before the next piperacillin/tazobactam infusion were not increased (median 0.2, IQR 0.2-0.35, P > 0.05), but 10% (3/30) were still > or = 0.5. With a cut-off level of > 0.7, no false-positive results were noted at any sampling time point.. We conclude that the clinical relevance of false-positive GM results during piperacillin/tazobactam treatment is small if samples are collected prior to infusion and if a cut-off level of > 0.7 is used.

Topics: Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Galactose; Humans; Mannans; Middle Aged; Penicillanic Acid; Piperacillin; Prospective Studies; Tazobactam

Topics: Antidiarrheals; Aspergillosis; False Positive Reactions; Female; Galactans; Galactose; Gastroesophageal Reflux; Humans; Infant; Infant Food; Infant Formula; Leukemia, Myeloid, Acute; Mannans; Plant Gums

Diagnosing invasive aspergillosis is difficult but might be improved by detection of circulating galactomannan. Although galactomannan antigenemia has been well studied in the detection of invasive aspergillosis in adult patients, little is known about the expression of circulating galactomannan in immunocompromised children with invasive aspergillosis.. We studied the expression of galactomannan antigen by enzyme immunoassay (EIA) in 990 serum samples from 56 pediatric oncology patients (ages 3 months to 18 years) of whom 17 had proven or probable invasive aspergillosis defined by the European Organization for Research and Treatment of Cancer-Mycoses Study Group criteria. Any sample with a galactomannan EIA Galactomannan index value of > or = 0.5 was considered positive.. At least 1 serum sample was positive for 11 of 17 pediatric oncology patients (65.7% sensitivity, 95% confidence interval: 38.3-85.7) with invasive aspergillosis. Galactomannan EIA was positive in 99 of 304 samples from patients with proven or probable invasive aspergillosis, and 7 of 686 (1.0%) samples from 39 control subjects resulted in a positive galactomannan EIA result. At least 1 sample tested positive in 5 of the 39 controls (12.8%, 95% confidence interval: 4.3-27.4). No significant association between accuracy and patient age was observed. Among the 7 evaluable galactomannan-positive patients with IA, the galactomannan EIA produced a positive result before clinical or radiographic evidence of infection in 6 cases, with a lead-time to diagnosis ranging from 1 day to 34 days (median: 10 days). In the remaining case, a positive galactomannan was observed on the same day as diagnosis by non-EIA methods.. The presence of circulating galactomannan is predictive of invasive aspergillosis in most pediatric oncology patients. Galactomannan antigenemia may precede clinical, microbiologic, or radiographic evidence of invasive aspergillosis.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Child; Child, Preschool; Galactose; Humans; Immunocompromised Host; Immunoenzyme Techniques; Infant; Mannans; Neoplasms; Predictive Value of Tests; Sensitivity and Specificity; Time Factors

Topics: Aged; Aged, 80 and over; Aspergillosis; Biomarkers; Female; Galactose; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Sensitivity and Specificity; Sputum

Detection of galactomannan antigen (GMA) in serum is the standard assay for the diagnosis of invasive aspergillosis (IA) in high-risk patients with hematological disorders. Detection of Aspergillus DNA in serum has been proposed, but its sensitivity is lower than that of GMA when small serum volumes (SSV) are used. In this study, we investigated whether extraction of DNA from large serum volumes (LSV) improves diagnostic yield. In a 13-month prospective study, we compared the performances of twice-weekly screening of serum for GMA by an enzyme immunoassay and weekly screening for Aspergillus fumigatus DNA by a real-time PCR (RT-PCR) assay of 1.0 ml (LSV) or 100 mul (SSV) of serum. We included 124 patients (138 treatment episodes), with 17 episodes of EORTC (European Organization for Research and Treatment of Cancer)/MSG (Mycoses Study Group)-documented IA. In all, 1,870 samples were screened for GMA. The sensitivity (Se), specificity (Sp), and positive and negative predictive values (PPV and NPV, respectively) of GMA for IA were 88.2%, 95.8%, 75%, and 98.3%, respectively. We screened 938 samples for Aspergillus DNA by using LSV; 404 of these samples were also tested with SSV. The Se, Sp, PPV, and NPV of RT-PCR were 100%, 96.7%, 81%, and 100%, respectively, with LSV and 76.5%, 96.7%, 81.3%, and 95.6%, respectively, with SSV. DNA detection gave a positive result when performed on LSV in two cases of IA where the GMA assay result remained negative. Furthermore, in four IA cases, DNA was detected earlier than GMA. The use of LSV for extraction improved the performance of the RT-PCR, which appears highly sensitive and specific for the early diagnosis of IA in high-risk patients with hematological disorders.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus fumigatus; DNA, Fungal; Early Diagnosis; Female; Galactose; Hematologic Diseases; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity; Serum; Time Factors

We have evaluated the Platelia Aspergillus enzyme immunoassay for detection of galactomannan in bronchoalveolar lavage (BAL) specimens in solid organ transplant patients with aspergillosis. The precision and reproducibility in serum or BAL to which galactomannan was added were similar. Sensitivity was 81.8% in patients with aspergillosis, and specificity was 95.8% in lung transplant patients who underwent BAL for surveillance for infection or rejection. Among transplant controls, positive results were more common in patients (i) who underwent diagnostic BAL performed for evaluation of symptoms or chest computed tomographic abnormalities, (ii) who had undergone lung transplantation, or (iii) who were colonized with Aspergillus. Galactomannan testing in BAL is useful for diagnosis of aspergillosis in transplant patients. The significance of positive results in patients without confirmed aspergillosis requires further evaluation.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Galactose; Humans; Immunoenzyme Techniques; Mannans; Reproducibility of Results; Sensitivity and Specificity

Serum galactomannan antigen detection was used for the diagnosis and follow-up of cardiac aspergillosis after surgery in 2 nonneutropenic patients. The galactomannan index, developed in response to surgical and antifungal therapies, could prove to be a valuable method for the diagnosis and follow-up of fungal infections in such patients.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Biomarkers; Child; Follow-Up Studies; Galactose; Heart Diseases; Humans; Male; Mannans

We studied 75 patients with non-hematological conditions from whom Aspergillus spp. were recovered from clinical specimens during the period March 2003 to August 2006. The patients were classified according to EORTC criteria and the presence of galactomannan (Platelia Aspergillus) in their sera was evaluated. Ten of these patients (13.3%) had proven or probable invasive aspergillosis, i.e., chronic obstructive pulmonary disease in five (50%), HIV infection in one (10%), lymphoma in one (10%), liver transplant in one (10%), solid malignancies in one (10%), and corticosteroid treatment in one (10%). The sensitivity, specificity, and positive and negative predictive values for the detection of galactomannan, using cut-offs of > or =0.5 ng/ml and > or =1 ng/ml were 60%/50%, 89.23%/100%, 46.15%/100%, and 93.55%/92.86% (p=0.001 and p<0.001), respectively. The determination of galactomannan in the sera of non-neutropenic patients could prove to be a useful microbiological finding when diagnosing invasive aspergillosis.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Galactose; Humans; Mannans; Prospective Studies

Invasive aspergillosis (IA) is an important cause of mortality in patients with hematologic malignancies. However, IA appears to be gaining a foothold in the intensive care unit (ICU) in patients without classical risk factors. A recent study described 89 cases of IA in patients in a medical ICU without leukemia or cancer. The diagnosis of IA remains difficult and is often established too late. Galactomannan (GM) is an exo-antigen released from Aspergillus hyphae while they invade host tissue.. This prospective single-center study was conducted to investigate the role of GM in bronchoalveolar lavage (BAL) fluid as a tool for early diagnosis of IA in the ICU.. All patients with risk factors identified in our earlier study were evaluated. BAL for culture and GM detection, serum GM levels, and computed tomography scan were obtained for all included patients with signs of pneumonia. Patients were classified as having proven, probable, or possible IA.. A total of 110 patients out of 1,109 admissions were eligible. There were 26 proven IA cases. Using a cutoff index of 0.5, the sensitivity and specificity of GM detection in BAL fluid was 88 and 87%, respectively. The sensitivity of serum GM was only 42%. In 11 of 26 proven cases, BAL culture and serum GM remained negative, whereas GM in BAL was positive.. IA is common in immunocompromised, critically ill patients. GM detection in BAL fluid seems to be useful in establishing or excluding the diagnosis of IA in the ICU.

Topics: Adult; Aged; Aspergillosis; Bronchoalveolar Lavage Fluid; Bronchoscopy; Cause of Death; Cross Infection; Diagnosis, Differential; Female; Galactose; Hospital Mortality; Humans; Intensive Care Units; Lung; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Mycological Typing Techniques; Opportunistic Infections; Pneumonia, Ventilator-Associated; Predictive Value of Tests; Prospective Studies; Survival Rate; Tomography, X-Ray Computed

We evaluated the impact of piperacillin/tazobactam (PT) therapy on galactomannan enzyme immunoassay (GEI) testing (Platelia; Bio-Rad, Marnes La Coquette, France). Galactomannan contents of PT batches were highly variable. We found that false-positive GEI results can be avoided by performance of blood sampling before PT administration and by using separate sites for blood sampling and for administration of antibiotics.

Topics: Adult; Antigens, Fungal; Aspergillosis; False Positive Reactions; Galactose; Humans; Immunoenzyme Techniques; Mannans; Penicillanic Acid; Piperacillin; Tazobactam

Topics: Aged; Antigens, Fungal; Aspergillosis; Aspergillus; False Positive Reactions; Fatal Outcome; Female; Galactose; Histoplasma; Histoplasmosis; Humans; Immunoenzyme Techniques; Mannans; Sputum

The diagnosis of invasive aspergillosis (IA) based on the detection of Aspergillus galactomannan (GM) is complicated by the presence of cross-reactive GM epitopes in patient specimens. We have developed a novel and specific Aspergillus antigen-capture enzyme-linked immunosorbent assay (ELISA) by the selection of two well-characterized monoclonal antibodies from 17 candidate antibodies. The epitopes recognized by the monoclonal antibodies were present on the cell walls of the hyphae and the conidia of Aspergillus species, which were circulating or excreted as immunodominant antigens during the acute phase of IA established in the animal models. The detection of experimental Aspergillus-mediated antigenemia was suitably sensitive, and the sensitivity was comparable to that of a commercial GM detection ELISA kit (the Platelia Aspergillus assay). Moreover, the specificity of this assay was 100% when it was used to test 382 serum specimens and 120 urine specimens from healthy individuals. Cross-reactivity with other common opportunistic fungi, such as Penicillium and Candida species, and with purified GM protein derived from Aspergillus was not evident. Therefore, the chemical nature of the epitopes captured in this assay is most likely not associated with the GM structure, indicating that this newly developed Aspergillus antigen-capture ELISA is a promising tool for the diagnosis of IA without the risk of the false-positive results that are problematic with current GM antigen assays.

Topics: Animals; Antibodies, Fungal; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus; Cell Wall; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Hyphae; Immunodominant Epitopes; Mannans; Mice; Mice, Inbred BALB C; Microscopy, Confocal; Microscopy, Fluorescence; Sensitivity and Specificity; Serum; Spores, Fungal; Urine

Although Aspergillus galactomannan (GM) antigen detection is widely applied in the diagnosis of invasive aspergillosis (IA), false-positive reactions with fungus-derived antibiotics, other fungal genera or the passage of dietary GM through injured mucosa are a matter of concern. The aim of this study was to investigate the cumulative incidence and risk factors for false-positive GM antigenaemia.. The records of 157 adult allogeneic haematopoietic stem cell transplantation (HSCT) recipients were retrospectively analysed. Episodes of positive GM antigenaemia, defined as two consecutive GM results with an optical density index above 0.6, were classified into true, false and inconclusive GM antigenaemia by reviewing the clinical course.. Twenty-five patients developed proven or probable IA with a 1 year cumulative incidence of 12.9%, whereas 50 experienced positive GM antigenaemia with an incidence of 32.2%. Among the total 58 positive episodes of the 50 patients, 29 were considered false-positive. The positive predictive value (PPV) was lower during the first 100 days than beyond 100 days after HSCT (37.5% versus 58.8%). Gastrointestinal chronic graft-versus-host disease (GVHD) was identified as the only independent significant factor for the increased incidence of false-positive GM antigenaemia (PPV 0% versus 66.7%, P = 0.02).. GM antigen results must be considered cautiously in conjunction with other diagnostic procedures including computed tomography scans, especially during the first 100 days after HSCT and in patients with gastrointestinal chronic GVHD.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; False Positive Reactions; Female; Follow-Up Studies; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Retrospective Studies

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Cross Infection; Galactose; Hospital Mortality; Intensive Care Units; Lung Diseases, Fungal; Mannans; Opportunistic Infections; Pneumonia, Ventilator-Associated; Predictive Value of Tests; Survival Rate

The aim of this study was to detect Aspergillus fumigatus-specific DNA by nested PCR (nPCR) in serum and bronchoalveolar lavage (BAL) specimens of experimentally infected rats and compare the results with (1-3)-beta-D-glucan (BDG) and galactomannan (GM) detection. Sixty Wistar rats, immunosuppressed with an intraperitoneal injection of cyclophosphamide (70 mg kg(-1)) were infected with 1 x 10(6)A. fumigatus conidia. The rats were killed on days 1, 3, 5, 7 and 9 postinfection in groups of six each and their BAL, blood and lungs were cultured. The A. fumigatus-specific DNA, BDG and GM in serum and BAL were detected by nPCR, Fungitell kit and Aspergillus Platelia kit respectively. Base line values were obtained by using sera from six healthy rats. Except the lungs, blood and BAL specimens of all the infected rats were negative for A. fumigatus culture. The BDG, GM and nPCR positivity in serum specimens was 80%, 77% and 63% respectively. The sensitivity of GM and nPCR tests in BAL specimens was 77% and 70% respectively. The data suggest that BDG and GM appear early in the course of infection, and have similar kinetics (r = 0.483, P = 0.007). Hence, their combined detection could be useful in the early diagnosis of invasive aspergillosis.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Bronchoalveolar Lavage Fluid; DNA, Fungal; Female; Galactose; Humans; Lung; Lung Diseases, Fungal; Mannans; Polymerase Chain Reaction; Rats; Rats, Wistar; Sensitivity and Specificity; Specific Pathogen-Free Organisms

Detection of circulating galactofuranose (galf) antigens, including galactomannan (GM), by the Platelia Aspergillus (PA) enzyme-linked immunosorbent assay (ELISA) is an important tool in the early diagnosis of invasive aspergillosis (IA). We used a modified pretreatment technique (MT) on consecutive negative PA ELISA plasma samples from IA patients in order to improve the detection of the fungal components present. Plasma samples (52) were collected from healthy donors, and 174 plasma samples with a galactomannan index (GMI) below 0.5 were collected from 25 unclassifiable and 23 IA patients. The PA ELISA reactivity of pretreated samples was determined before (conventional technique [CT]) and after (MT) filtration using a Microcon filter with a 50-kDa cutoff (Millipore). For the MT, the sensitivity of the PA ELISA increased from 42.9% (CT) to 78.6% (MT) using a cutoff for the GMI of 1.5 in the probable and proven group, whereas specificity slightly decreased from 98.7% to 96.1% in the control group. The 10-fold concentration step increased the GMI as high as 121-fold. The MT resulted not only in positive reactivity in samples that tested negative with the CT but also in the earlier detection of antigen by 2 to 17 days.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Child; Child, Preschool; Edetic Acid; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Sensitivity and Specificity

A reliable diagnosis of invasive aspergillosis (IA) is hampered by the difficulty in obtaining suitable tissue samples. To evaluate the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the LightCycler PCR for the diagnosis of IA, 536 blood samples were collected over a 22-month period from 62 paediatric patients (median age 10 years, range 1-18) considered at risk of IA. The galactomannan antigen (GM) and fungal DNA were assessed on serial blood samples. IA was diagnosed in eight of 62 patients (13%): proven, five, probable, three. Sensitivity, specificity, PPV and NPV of LightCycler PCR varied according to the number of positive samples used to define positivity: 88%; 37%; 17% and 95% for single sample positivity; and 63%, 81%, 33% and 94% for serial sample positivity respectively. The concordance between positivity of LightCycler PCR assay and the diagnosis of IA was 79%. The single positivity of LightCycler PCR assay showed a good sensitivity for the diagnosis of IA in paediatric patients. The high NPV makes LightCycler PCR a promising tool in addition to GM testing to design a strategy of pre-emptive antifungal therapy, although further validation studies are needed.

Topics: Adolescent; Aspergillosis; Child; Child, Preschool; DNA, Fungal; Female; Galactose; Hematologic Neoplasms; Humans; Infant; Male; Mannans; Polymerase Chain Reaction; Predictive Value of Tests; Sensitivity and Specificity

PREMISES AND OBJECTIVES: Timely diagnosis is of critical importance for the prognosis of invasive aspergilosis (IA) patients. Over recent years, IA detection of galactomannan using the ELISA method has assumed growing importance. The objective of the study was to analyse the usability of the method in current clinical practice of a hemato-oncological ward.. From May 2003 to October 2006, blood samples were taken from patients at IA risk to detect galactomannan (GM) in serum using the ELISA method. The patients who underwent the tests were classified by the probability of IA presence on the basis of the results of conventional diagnostic methods and section findings.. A total of 11,360 serum samples from 911 adult patients were tested for GM presence. IA (probable/proven) was diagnosed in 42 (4.6%) of them. The rates of sensitivity, specificity, positive and negative predictive value of galactomannan detection for IA diagnosis in our ward were, respectively, 95.2%, 90.0%, 31.5% and 99.7%. The principal causes of the limited positive predictive value of the test were the high percentage of false-positive test results (mainly caused by concomitant administration of some penicillin antibiotics or Plasma-Lyte infusion solution), as well as the fact that a large percentage of patients we examined fell within the group of patients with hematological malignity with a very low prevalence of IA.. GM detection in serum is associated with high sensitivity and excellent negative predictive value in IA diagnosis in hemato-oncological patients. Knowledge and elimination of possible causes of false-positive results as well as focusing the screening on patients at greatest risk of infection are necessary for an even better exploitation of the test.

Topics: Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Enzyme-Linked Immunosorbent Assay; Galactose; Hematologic Neoplasms; Humans; Mannans; Opportunistic Infections; Predictive Value of Tests; Sensitivity and Specificity

Galactomannan is an Aspergillus-specific polysaccharide released during aspergillosis and is detected by the quantitative serum galactomannan index (GMI) test. Preclinical and preliminary clinical reports have suggested a good correlation between GMI and aspergillosis outcome.. We reviewed the literature to assess the strength of correlation between GMI and aspergillosis outcome using the kappa correlation coefficient. We included 27 studies that enrolled patients with hematological cancer and proven or probable aspergillosis and that used sequential GMI testing. We examined the 3 following outcomes: survival (survival vs. death), global outcome (survival vs. death [including autopsy findings]), and autopsy outcome (autopsy findings only).. Overall, 257 patients fulfilled criteria for proven or probable aspergillosis and were eligible for outcome evaluation. Correlation between GMI (within

Topics: Aspergillosis; Aspergillus; Galactose; Hematologic Neoplasms; Humans; Mannans; Meta-Analysis as Topic; Prognosis; Survival Analysis; Treatment Outcome

Topics: Aspergillosis; Aspergillus; Galactose; Humans; Mannans; Treatment Outcome

The Aspergillus galactomannan test is a valuable tool in the diagnosis of invasive aspergillosis. We hereby report a high rate of false-positive results by the Platelia Aspergillus galactomannan antigen test (Bio-Rad Laboratories) for patients treated with amoxicillin-clavulanate.

Topics: Amoxicillin-Potassium Clavulanate Combination; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Galactose; Humans; Mannans

Early diagnosis of invasive pulmonary aspergillosis is problematic in some patient groups due to the lack of rapid, sensitive, specific, and reliable diagnostic tests. Fungal burden and therapeutic efficacy were assessed by survival, quantitative culture (CFU counts), galactomannan enzyme immunoassay (GM-EIA), and quantitative PCR (qPCR) in a new guinea pig model of invasive pulmonary aspergillosis using an aerosol challenge. At 1 day postinfection, qPCR determined that the pulmonary fungal burden was 2 log(10) higher than that determined by CFU counting and increased significantly (P < 0.03) over time. In contrast, the tissue burden assessed by CFU counting did not rise over the course of the study. Therapy with the antifungal drug voriconazole produced statistically significant decreases in pulmonary fungal burden, as detected by CFU counting (P < 0.02), qPCR, and GM-EIA (both P < 0.0002). Daily assessment of the progression of fungal infection in serum was performed by qPCR and GM-EIA. GM-EIA demonstrated a statistically significant reduction in the fungal load on days 6 and 7 in voriconazole-treated animals compared to time-matched controls (P < 0.02). Confirmation of fungal tissue burden by two or more methods should provide a more precise account of the burden, allowing improved assessment of diagnostic and therapeutic strategies in invasive pulmonary aspergillosis.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Base Sequence; Colony Count, Microbial; Disease Models, Animal; DNA Primers; DNA, Fungal; Galactose; Guinea Pigs; Humans; Immunoenzyme Techniques; Lung; Lung Diseases, Fungal; Male; Mannans; Mycology; Polymerase Chain Reaction; Pyrimidines; Triazoles; Voriconazole

The aim of this study was to determine whether interleukin-1 alpha (IL1alpha), interleukin-1 beta (IL1beta), and IL1 receptor antagonist (IL1Ra) polymorphisms are implicated in invasive pulmonary aspergillosis (IPA) pathogenesis.. Subjects comprised 110 hematological patients and 148 healthy controls. Genotypic and allelic frequencies were similar between hematological patients and controls. IPA was diagnosed in 59 of the 110 patients according to consensus criteria published by the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group (EORTC/IFICG).. Individual locus analysis showed that IL1alpha and IL1Ra polymorphisms were not associated with the presence of IPA (p = 0.560 and p = 0.680, respectively). However, a trend towards a higher presence of IL1beta( - ) (511TT) genotype (or IL1beta(-511T) allele) in the IPA group than in the non-IPA patient group (p = 0.092 and p = 0.095, respectively) was found. Haplotype analysis revealed that VNTR2/-889C/-511T haplotype was strongly associated with susceptibility to develop IPA infection (p = 0.020). Haplotype analysis also showed an association between VNTR2/-889C/-511C haplotype and resistance to IPA infection (p = 0.028). Furthermore, patients with IL1Ra VNTR2/2 and IL1beta(-511)T/T genotypes had a higher positive serum galactomannan percentage versus patients with other genotypes. Finally, C-reactive protein (CRP) production was significantly associated with IL1 gene cluster polymorphisms, although CRP values were similar between IPA and non-IPA groups.. These findings indicate a critical role of IL1 gene cluster polymorphisms in the susceptibility to IPA infection and CRP production.

Topics: Aspergillosis; Aspergillus; C-Reactive Protein; Drug-Related Side Effects and Adverse Reactions; Female; Galactose; Genetic Predisposition to Disease; Haplotypes; Hematologic Neoplasms; Humans; Immunocompromised Host; Interleukin-1alpha; Interleukin-1beta; Lung Diseases, Fungal; Male; Mannans; Multigene Family; Polymorphism, Single Nucleotide; Receptors, Interleukin-1 Type I; Statistics as Topic

The occurrence of invasive fungal infection (IFIs) in a pediatric hematology/oncology unit after renovation of the ventilation system, and initiating routine azole antifungal prophylaxis was monitored. In addition, the value of serial screening for Aspergillus galactomannan (GM) for diagnosing invasive aspergillosis was assessed.. A total of 98 consecutive high-risk pediatric patients were prospectively surveyed for signs of IFI and weekly monitored for serum GM. The data was not made available to treating physicians.. Only 2 patients had proven and 27 possible IFI based on the European Organization for Research and Treatment of Cancer/Mycoses Study Group definitions. The incidence of proven IFI was 1/31 (3.2%) in the allogeneic stem cell transplant (SCT) (Aspergillus spp), 0/26 in the autologous SCT, and 1/60 (1.6%) in the induction therapy group (C. krusei). GM was detected at least in one tested sample in 12/98 patients (12.2%), in five patients in two or more sequential samples. In the latter group, IFI was proven in one patient and could not be excluded in the others. Four of the five patients belonged to the 31 allogeneic and one to the 26 autologous SCT patients. In patients with only one positive GM test none developed signs of IFI and only one received empirical amphotericin B.. With the currently used preventative and prophylactic measures, IFI is uncommon in children with high-risk for infection. Regular screening for GM could be useful among allogeneic SCT patients and two positive samples should prompt further investigative procedures and pre-emptive antifungal therapy.

Topics: Adolescent; Antifungal Agents; Antineoplastic Combined Chemotherapy Protocols; Aspergillosis; Child; Child, Preschool; Female; Galactose; Humans; Infant; Leukemia, Myeloid, Acute; Male; Mannans; Monitoring, Physiologic; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Remission Induction; Retrospective Studies; Risk Factors; Stem Cell Transplantation; Transplantation, Autologous; Transplantation, Homologous

The purpose of this study was to assess the antifungal activity, pharmacokinetics, and tissue distribution of amphotericin B (AmpB) following the administration of Abelcet and AmBisome alone and in combination with Caspofungin to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (2.1-2.5 x 10(7) colony forming units [CFU]) was injected via the jugular vein; 48 h later male albino Sprague-Dawley rats (350-400 g) were administered either a single intravenous (i.v.) dose of Abelcet (5 mg AmpB/kg; n = 6), AmBisome (5 mg AmpB/kg; n = 6), Caspofungin (3 mg/kg; n = 5), Abelcet (5 mg AmpB/kg) plus Caspofungin (3 mg/kg) (n = 6), AmBisome (5 mg AmpB/kg) plus Caspofungin (3 mg/kg) (n = 7), or physiologic saline (non-treated controls; n = 6) once daily for 4 days. Antifungal activity was assessed by organ CFU concentrations and plasma galactomannan levels. Plasma and tissue samples were taken from each animal for AmpB pharmacokinetic analysis and tissue distribution determinations. Abelcet treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 73% compared to non-treated controls. Ambisome treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 69% compared to non-treated controls. Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 80% compared to non-treated controls. Abelcet plus Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 81% compared to non-treated controls. Ambisome plus Caspofungin treatment significantly decreased total fungal CFU concentrations recovered in all the organs added together by 98% compared to non-treated controls. Abelcet treatment significantly decreased plasma galactomannan levels by 50 and 75% 96 h following the initiation of treatment in the absence and presence of Caspofungin co-therapy, respectively. AmBisome treatment significantly decreased plasma galactomannan levels by 73 and 78% 96 h following the initiation of treatment in the absence and presence of Caspofungin co-therapy, respectively. Co-administration of Caspofungin with Abelcet and AmBisome did not significantly alter the plasma concentration-time profile, pharmacokinetic parameters, and tissue distribution of AmpB. Taken together, our findings suggest that an alternative mechanism, possibly at t

Topics: Amphotericin B; Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Caspofungin; Colony Count, Microbial; Disease Models, Animal; Drug Administration Schedule; Drug Combinations; Drug Therapy, Combination; Echinocandins; Galactose; Injections, Intravenous; Lipopeptides; Male; Mannans; Peptides, Cyclic; Phosphatidylcholines; Phosphatidylglycerols; Rats; Rats, Sprague-Dawley; Tissue Distribution

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Electrolytes; False Positive Reactions; Galactose; Humans; Mannans; Plasma Substitutes

A case of cerebral aspergillosis was diagnosed by the detection of Aspergillus flavus-specific DNA in brain biopsy and serum specimens. The diagnosis was also supported by detection of elevated levels of galactomannan and (1-->3)-beta-d-glucan in serum specimens. Despite the presence of dichotomously branched septate hyphae in brain biopsy, the culture remained negative. The inability to isolate the organism in culture suggested that combined therapy of AmBisome and caspofungin was fungicidal for the fungus in the brain abscess.

Topics: Amphotericin B; Antifungal Agents; Aspergillosis; Aspergillus flavus; beta-Glucans; Brain; Brain Diseases; Caspofungin; Diagnosis, Differential; DNA, Fungal; Echinocandins; Electrophoresis, Agar Gel; Galactose; Humans; Lipopeptides; Male; Mannans; Middle Aged; Peptides, Cyclic; Polymerase Chain Reaction

Little is known about the pathogenesis of invasive pulmonary aspergillosis and the relationship between the kinetics of diagnostic markers and the outcome of antifungal therapy.. An in vitro model of the human alveolus, consisting of a bilayer of human alveolar epithelial and endothelial cells, was developed. An Aspergillus fumigatus strain expressing green fluorescent protein was used. Invasion of the cell bilayer was studied using confocal and electron microscopy. The kinetics of culture, polymerase chain reaction, and galactomannan were determined. Galactomannan was used to measure the antifungal effect of macrophages and amphotericin B. A mathematical model was developed, and results were bridged to humans.. A. fumigatus penetrated the cellular bilayer 14-16 h after inoculation. Galactomannan levels were inextricably tied to fungal invasion and were a robust measure of the antifungal effect of macrophages and amphotericin B. Neither amphotericin nor macrophages alone was able to suppress the growth of A. fumigatus; rather, the combination was required. Monte Carlo simulations showed that human dosages of amphotericin B of at least 0.6 mg/kg were required to achieve adequate drug exposure.. This model provides a strategy by which relationships among pathogenesis, immunological effectors, and antifungal drug therapy for invasive pulmonary aspergillosis may be further understood.

Topics: Amphotericin B; Antifungal Agents; Arteries; Aspergillosis; Aspergillus fumigatus; Cell Line, Tumor; Endothelial Cells; Galactose; Humans; In Vitro Techniques; Kinetics; Lung; Lung Diseases, Fungal; Macrophages; Mannans; Models, Biological; Monte Carlo Method

Topics: Aspergillosis; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Galactose; Humans; Mannans; Neoplasms

Topics: Adult; Antifungal Agents; Antigens, Fungal; Aspergillosis; beta-Glucans; Caspofungin; Echinocandins; Fungal Proteins; Galactose; Humans; Lipopeptides; Mannans; Peptides, Cyclic

Invasive aspergillosis is difficult to diagnose in patients undergoing hematopoietic stem cell transplantation (HSCT). In 2003, a serum enzyme-linked immunosorbent assay (ELISA) test for the detection of galactomannan (a glycoprotein found on the Aspergillus cell wall) became available in the United States. In 2004, patients undergoing HSCT were screened biweekly with the galactomannan ELISA at our institution. We performed a retrospective chart review of 121 SCT patients who underwent galactomannan testing. Thirteen of the patients (10.7%) had at least 1 positive galactomannan ELISA, and 4 had multiple positive tests. When calculated in reference to a proved or probable diagnosis of aspergillosis, the galactomannan ELISA had a sensitivity of 0.50 and a specificity of 0.94. The positive predictive value was 0.46, and the negative predictive value was 0.94. Galactomannan ELISA had fewer false-positive and false-negative results in pediatric patients than in adult patients. In 4 of the 12 cases of invasive aspergillosis, the galactomannan ELISA was positive before other microbiologic evidence of aspergillosis was available. In these cases, a positive ELISA predated culture and cytologic evidence of invasive aspergillosis by a mean of 5 days (range, 1-8 days). Our findings indicate that a biweekly serum galactomannan ELISA is a highly specific diagnostic tool for detecting invasive aspergillosis in patients undergoing HSCT. When used regularly, the ELISA may allow for earlier diagnosis of invasive aspergillosis in some patients. The test is troubled by a low sensitivity and high frequency of false-negative tests.

Topics: Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; False Negative Reactions; False Positive Reactions; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Retrospective Studies; Sensitivity and Specificity

Topics: Aged; Aspergillosis; False Positive Reactions; Female; Galactose; Gluconates; Humans; Infusions, Intravenous; Mannans; Solutions

This present study was undertaken to examine the role of the host response to Aspergillus fumigatus in the development of clinical symptoms of invasive pulmonary aspergillosis (IPA). The natural outcome and response to IPA infection varies between individuals. Whereas some variation may be attributable to fungi and environmental variables, it is probable that host genetic background also plays a significant role. Interleukin (IL)-10 has a key function in the regulation of cellular immune responses and is involved in various inflammatory diseases. IL-10 promoter carries a polymorphism that has been associated to production levels. Our aim was to investigate the role of this polymorphism in susceptibility to develop IPA infection. The study included 120 haematological patients and 124 age and sex-matched controls and bi-allelic IL-10 -1082(G/A) polymorphism was examined. Genotypic (p=0.385) and allelic frequencies (p=0.527, OR=0.89, 95% CI=0.78-1.60) were similar between patients and healthy controls. IPA was diagnosed in 59 of the 120 patients according to consensus criteria published by the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group (EORTC/IFICG). Our results provide evidence that IL-10 -1082(AA) genotype is associated with resistance to develop IPA (p=0.001). Allele frequency of IL-10 -1082A allele was weakly associated with susceptibility to develop IPA infection (p=0.052). In conclusion, these results suggest that differential production of IL-10 may alter the risk for IPA in haematological patients.

Topics: Adult; Aspergillosis; Female; Galactose; Genetic Predisposition to Disease; Humans; Interleukin-10; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Polymorphism, Restriction Fragment Length; Promoter Regions, Genetic; Prospective Studies

We review the experience at our institution with galactomannan (GM) testing of bronchoalveolar lavage (BAL) fluid in the diagnosis of invasive pulmonary aspergillosis (IPA) among solid-organ transplant recipients. Among 81 patients for whom BAL GM testing was ordered (heart, 24; kidney, 22; liver, 19; lung, 16), there were five cases of proven or probable IPA. All five patients had BAL GM of > or = 2.1 and survived following antifungal therapy. The sensitivity, specificity, and positive and negative predictive values for BAL GM testing at a cutoff of > or = 1.0 were 100%, 90.8%, 41.7%, and 100%, respectively. The sensitivity of BAL GM testing was better than that of conventional tests such as serum GM or BAL cytology and culture. Moreover, a positive BAL GM test diagnosed IPA several days to 4 weeks before other methods for three patients. Twelve patients had BAL GM of > or = 0.5 but no evidence of IPA. Among these, lung transplant recipients accounted for 41.7% (5/12) of the false-positive results, reflecting frequent colonization of airways in this population. Excluding lung transplants, the specificity and positive predictive value for other solid-organ transplants increased to 92.9% and 62.5%, respectively (cutoff, > or = 1.0). In conclusion, BAL GM testing facilitated more-rapid diagnoses of IPA and the institution of antifungal therapy among non-lung solid-organ transplant recipients and helped to rule out IPA.

Topics: Adolescent; Adult; Aged; Aspergillosis; Bronchoalveolar Lavage Fluid; Child; Child, Preschool; Female; Galactose; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Organ Transplantation; Predictive Value of Tests; Sensitivity and Specificity

Many health care centers worldwide use the Platelia Aspergillus enzyme immunoassay (PA-EIA; Bio-Rad Laboratories) for diagnosis of invasive aspergillosis (IA). A cutoff optical density (OD) index of 1.5 was originally recommended by the manufacturer, but in practice, most institutions use lower cutoff values. Moreover, a cutoff OD index of 0.5 was recently approved in the United States. In the present study, we set out to optimize the cutoff level by performing a retrospective analysis of PA-EIA values for samples that had been obtained prospectively from adult patients at risk for IA at 2 European health care centers.. In total, 239 treatment episodes were included of which there were 19 episodes of proven IA and 19 episodes of probable IA. Per-episode and per-test analyses and receiver operating characteristic curves were used to determine the optimal cutoff value.. In the per-episode analysis, lowering the cutoff OD index for positivity from 1.5 to 0.5 increased the overall sensitivity by 21% (from 76.3% to 97.4%) but decreased the overall specificity by 7% (from 97.5% to 90.5%). Requiring 2 consecutive samples with an OD index > or = 0.5 resulted in the highest test accuracy, with an improved positive predictive value. At a cutoff OD index of 0.5, the antigen test result was positive during the week before conventional diagnosis in 65% of cases and during the week of diagnosis in 79.5% of cases.. A cutoff OD index of 0.5--identical to the approved cutoff in the United States--improves the overall performance of the PA-EIA for adult hematology patients.

Topics: Adolescent; Adult; Aged; Aspergillosis; Aspergillus; Female; Galactose; Humans; Immunoenzyme Techniques; Male; Mannans; Middle Aged; Neutropenia; Retrospective Studies; ROC Curve; Sensitivity and Specificity

This study investigated the diagnostic value of Platelia Aspergillus enzyme immunoassay (EIA) for galactomannan (GM) antigen in patients at risk of invasive aspergillosis (IA), and its association with clinical course and outcome.. A total of 304 blood samples were collected from 189 patients at risk of IA during a 1-year period at a tertiary referral center. Classification of IA was made on the basis of the European Organization for Research and Treatment of Cancer case definitions.. Of the 189 patients, 5 had proven IA, 9 had probable IA, 26 had possible IA, and 149 had no IA. Diagnostic levels of GM were detected in 80% of proven and in 77% of probable IA cases. The overall sensitivity, specificity, and positive and negative predictive values for this assay, using a 1.5 index cut-off value, were 78.6%, 93.9%, 55.0%, and 97.9%, respectively. With the 0.5 index cut-off value, the sensitivity would increase to 100%. A close relationship was found between clinical course and the kinetics of GM indices in survivors.. The Platelia Aspergillus EIA is a useful screening test for the detection of IA. Regular monitoring of the kinetics of GM-EIA indices is a useful predictor of clinical course and outcome.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Child; Female; Galactose; Humans; Immunoenzyme Techniques; Kinetics; Male; Mannans; Middle Aged; Predictive Value of Tests; Sensitivity and Specificity; Time Factors

Topics: Aspergillosis; Equipment Contamination; False Positive Reactions; Galactose; Humans; Immunoenzyme Techniques; Mannans; Paper

The clinical utility of Platelia Aspergillus enzyme immunoassay (EIA) for galactomannan (GM) antigen detection in bronchoalveolar lavage (BAL) for the diagnosis of invasive aspergillosis (IA) in lung transplant recipients is not known.. BAL fluid samples from consecutive lung transplant recipients who underwent bronchoscopy were prospectively analyzed for GM.. A total of 333 BAL samples from 116 patients were tested. Invasive aspergillosis was documented in 5.2% (6/116) of the patients. Samples analyzed included 9 BALs from two patients with proven IA, 19 BALs from four patients with probable IA, and 305 BALs from 110 patients without IA. At the index cutoff value of > or =0.5, the sensitivity was 60%; specificity was 95%, with positive and negative likelihood ratios of 14 and 0.41, respectively. Increasing the index cutoff value to > or =1.0 yielded a sensitivity of 60%, a specificity of 98%, and the positive and negative likelihood ratios of 28 and 0.40, respectively. Two of six patients with IA receiving antifungal prophylaxis had false-negative results.. A Platelia EIA index cut-off > or =1.0 in the BAL fluid in a lung transplant recipient with a compatible clinical illness may be considered as suggestive of IA.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Aspergillosis; Bronchoalveolar Lavage Fluid; Drug Therapy, Combination; Female; Galactose; Humans; Immunosuppressive Agents; Lung Transplantation; Male; Mannans; Middle Aged; Postoperative Complications; ROC Curve

Assessing the outcome of patients with invasive pulmonary aspergillosis by using conventional criteria is difficult, particularly when clinical and radiologic worsening coincides with neutrophil recovery. Usually, it is assumed that this deterioration is related to progressive aspergillosis, prompting changes in patient management. However, its temporal relation with neutrophil recovery suggests that it may be caused by an immune reconstitution syndrome (IRIS). Galactomannan is an Aspergillus-specific polysaccharide that is released during aspergillosis and is detected by the serum galactomannan test, which has been approved by the United States Food and Drug Administration for the diagnosis of invasive aspergillosis. In this study, the authors used sequential galactomannan testing to distinguish IRIS responses from progressive aspergillosis.. From April 2001 to December 2006, patients with hematologic malignancies underwent galactomannan screening during periods when they were at risk. The clinical and laboratory findings from patients who had >or=2 consecutive positive galactomannan assays (optical density, >or=0.5) were reviewed.. Nineteen neutropenic patients with aspergillosis developed clinical and radiologic pulmonary deterioration during neutrophil recovery. Deterioration coincided with microbiologic response, as documented by rapid normalization of serum galactomannan, and, in 16 patients, was followed by complete clinical response and survival at 3 months, although there were no changes in antifungal therapy. The 3 patients who died during the first month had no evidence of aspergillosis at autopsy examination.. The authors propose that IRIS was responsible for the current findings and provide a definition for the syndrome. They also recommend serial galactomannan testing to guide aspergillosis management. Declining galactomannan values imply IRIS with an aspergillus response and obviate the need for invasive procedures and alternative antifungal therapies, whereas persistent galactomannan elevation indicates progressive aspergillosis and requires prompt treatment modification.

Topics: Acquired Immunodeficiency Syndrome; Adolescent; Adult; Aged; Aspergillosis; Galactose; Humans; Lung; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Neoplasms; Neutropenia

The aim of this study was to evaluate the diagnostic value of Aspergillus terreus-specific DNA, (1-3)-beta-d-glucan (BDG), and galactomannan (GM) in immunosuppressed mice infected intravenously with A. terreus conidia and sacrificed in groups of 12 each on days 1, 3, 5, 7, and 9. A. terreus-specific DNA, BDG, and GM in serum and bronchoalveolar lavage (BAL) were detected by nested polymerase chain reaction (nPCR), Fungitell kit (Associates of Cape Cod, E. Falmouth, MA), and Aspergillus Platelia kit (Bio-Rad, Marnes-laCoquette, France), respectively. Cultures of lung homogenate of all the animals yielded A. terreus. The BDG positivity, GM positivity, and nPCR positivity in serum specimens were 43%, 78%, and 73%, respectively. Combined detection enhanced the positivity to 95% for A. terreus DNA and GM, 83% for GM and BDG, and 95% for DNA, GM, and BDG. In BAL, the GM positivity and nPCR positivity were 80% and 81%, respectively, whereas combined detection increased the positivity to 98%. Detection of GM and DNA offers a sensitive and specific diagnostic option for invasive aspergillosis.

Topics: Animals; Aspergillosis; Aspergillus; beta-Glucans; Bronchoalveolar Lavage Fluid; DNA, Fungal; Female; Galactose; Humans; Mannans; Mice; Mice, Inbred BALB C; Polymerase Chain Reaction; Reagent Kits, Diagnostic; Sensitivity and Specificity; Specific Pathogen-Free Organisms

The galactomannan (GM) assay is an approved noninvasive test for detection of invasive aspergillosis (IA) that has been validated in adult patients with hematologic malignancies who are undergoing bone marrow transplantation. There have been few studies with this assay in pediatric patients, but early reports suggest that there may be differences in the performance such that false-positive GM tests in pediatric patients are more common than in adult patients.. We performed a prospective study in pediatric hematopoietic stem cell transplant recipients with twice-weekly sampling for GM detection during the highest risk periods of neutropenia and graft-versus-host disease. We analyzed 826 serum samples from 64 patients, including 15 serum samples from one patient diagnosed with probable IA according to defined criteria.. Twenty of 811 samples tested positive on repeat testing (specificity, 97.5%; 95% CI: 96.2-98.4%) including samples from 8 of 63 patients without clinical evidence of IA according to study criteria (specificity, 87.3%; 95% CI: 76.9-93.4%). Eleven patients received piperacillin/tazobactam therapy, and 4 of the 11 patients had a positive assay result coinciding with the dates of piperacillin/tazobactam administration. When samples from these patients were excluded, specificity increased to 98.4% (95% CI: 97.2-99.1%) by sample and to 91.5% (95% CI: 81.6-96.3%) by patient.. The GM assay holds promise for early, noninvasive diagnosis of IA in high-risk children and false-positive results were not common or unexplainable. This study supports further validation of this assay in a large-scale, pediatric-dedicated format.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Child; Enzyme-Linked Immunosorbent Assay; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Mannans; Predictive Value of Tests; Sensitivity and Specificity

In this prospective study including 78 adult patients with haematological malignancy (90 episodes) we performed galactomannan (GM) (Platelia Aspergillus) screening twice weekly for the diagnosis of invasive aspergillosis. There were five proven and four probable invasive aspergillosis cases. The sensitivity, specificity and positive and negative predictive values were 100, 88, 47 and 100%, respectively. There were eight patients with false positive GM (10.2%). In six patients the false GM reactivity was due to the administration of piperacillin-tazobactam (P-T). A significant association was found between false positive GM (= or > 0.5) and the administration of P-T (p < 0.01). Two other patients with no invasive aspergillosis (2.5%) and false GM reactivity had graft versus host disease (GVHD) and one of them had also mucositis grade IV. The kinetic patterns of false positive GM due to P-T is discussed.

Topics: Adolescent; Adult; Aged; Anti-Bacterial Agents; Antineoplastic Combined Chemotherapy Protocols; Artifacts; Aspergillosis; Biomarkers; Combined Modality Therapy; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Fungemia; Galactose; Graft vs Host Disease; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Mucositis; Neutropenia; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity

Determining the outcome of patients with aspergillosis can be particularly difficult because patients with aspergillosis are at risk for other conditions that mimic this infection. Galactomannan is an Aspergillus-specific antigen released during invasive aspergillosis and is detected by the quantitative serum galactomannan index (GMI) test.. Using a kappa correlation coefficient test (KCC), the strength of correlation was determined between GMI and survival outcome of aspergillosis among 56 adults with hematologic cancer (90% had myeloma) who underwent serial GMI monitoring until hospital discharge or death.. All 56 patients received antineoplastic therapy (myeloablative followed by stem cell transplantation [autologous in 21 patients and allogeneic in 3 patients] or nonmyeloablative therapy [32 patients]). The overall correlation between survival outcome and GMI was excellent (KCC = 0.8609; 95% confidence interval [95% CI], 0.7093-1.000 [P < .0001]) and was comparable among neutropenic and nonneutropenic patients (KCC = 0.8271; 95% CI, 0.6407-1.000 [P < .0001] and KCC = 1.0; 95% CI, 1-1 [P = .0083], respectively).. The survival outcome of patients with aspergillosis strongly correlated with serum GMI. These findings have important implications for patient care and clinical trials of mold-active antifungal agents.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Combined Modality Therapy; Dexamethasone; Female; Galactose; Glucocorticoids; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Multiple Myeloma; Predictive Value of Tests; Prognosis; Stem Cell Transplantation; Survival Analysis

We compared the sensitivity and specificity of galactomannan enzyme-linked immunosorbent assay (GM-ELISA) with that of a newly developed version of the commercially available latex agglutination test (GM-LATEX) in our patient population. Serum samples were collected from 144 healthy adult donors (controls); another 17 consecutive hematologic malignancy (HM) patients with probable or proven invasive aspergillosis (IA) were tested. Clinical, microbiological, and pathological data were obtained. The sensitivity of the GM-LATEX test was 53% per patient and 66% per sample and that of the GM-ELISA test was 41% per patient and 21% per sample. Both tests demonstrated a specificity of 99%. The GM-LATEX test demonstrated a superior sensitivity in terms of detecting galactomannan and thus may be useful in the diagnosis of IA in patients with HM.

Topics: Adult; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Hematologic Neoplasms; Humans; Latex Fixation Tests; Mannans; Sensitivity and Specificity

To evaluate the value of serum galactomannan (GM) detection for diagnosis of invasive aspergillosis (IA) in hematopoietic stem cell transplant recipients.. The serum GM concentration in 167 sera from 46 patients was detected by Platelia Aspergillus double-sandwich enzyme linked immunosorbent assay (PADSELISA). According to the diagnostic criteria of invasive fungal infections in China, the diagnostic changes were evaluated, the sensitivity, specificity and predictive values were calculated.. The sensitivity and specificity of the PADSELISA were 81.8% and 93.3% and the positive and negative predictive values were 90.0% and 87.5% respectively. There were 15 positive cases, and 31 negative cases, and the probable IA cases were increased from 11 to 19 after the GM detection. Moreover, the serous level of galactomannan was correlated with the prognosis of the IA.. The PADSELISA for GM detection is a reliable method for early diagnosis and treatment of IA in hematopoietic stem cell transplant recipients.

Topics: Adult; Aspergillosis; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Postoperative Complications; Sensitivity and Specificity

The purpose of this study was to assess the antifungal activity of a new oral amphotericin B (AmpB) lipid-based formulation following administration to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (2.1-2.5 x 10(7) colony forming units [CFU]) were injected via the jugular vein; 48h later male albino Sprague-Dawley rats (350-400 g) were administered either a single oral dose of AmpB incorporated into Peceol (50 mg AmpB/kg), physiologic saline (nontreated controls) or Peceol alone (vehicle control) once daily for 4 days. To assess antifungal activity Brain, Lung, Heart, Liver, Spleen and Kidney sections were homogenized with normal saline (1 mL/g of tissue) and a 0.1-mL aliquot was spread plated onto a Sabourand dextrose agar plate. The plates were incubated for 48 hr at 37 degrees C, at which time the number of fungal CFU were determined and corrected for tissue weight. In addition, plasma galactomannan antigen concentrations were determined. Data was reported as mean +/- standard error of the mean. The AmpB-Peceol oral formulation significantly decreased total fungal CFU concentrations recovered in all the organs added together, brain CFU concentrations, spleen CFU concentrations and plasma galactomannan antigen concentrations compared to baseline. No significant differences in lung, heart, liver and kidney CFU concentrations between treatment and control groups were observed. Peceol vehicle control did not exhibit any antifungal activity. These findings suggest that a new oral lipid-based formulation of AmpB incorporated into Peceol can significantly decrease brain and spleen CFU concentrations and plasma galactomannan antigen concentrations compared to non-treated controls.

Topics: Administration, Oral; Amphotericin B; Animals; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Brain; Galactose; Intestinal Absorption; Male; Mannans; Oleic Acids; Rats; Rats, Sprague-Dawley; Spleen; Stem Cells

The Aspergillus galactomannan enzyme-linked immunosorbant assay (EIA) has been demonstrated to facilitate rapid and sensitive detection of invasive aspergillosis. However, test specificity has not been fully evaluated in non-Aspergillus fungal species. Of 53 fungal isolates, cross-reactivity was observed with 5 non-Aspergillus spp.: Blastomyces dermatitidis, Nigrospora oryzae, Paecilomyces lilacinus, Penicillium chrysogenum, and Trichothecium roseum.

Topics: Aspergillosis; Aspergillus; Cross Reactions; Galactose; Humans; Immunoenzyme Techniques; Mannans; Mycological Typing Techniques; Mycoses; Sensitivity and Specificity

Topics: Aspergillosis; False Positive Reactions; Galactose; Gluconates; Humans; Magnesium Chloride; Mannans; Potassium Chloride; Serum; Sodium Acetate; Sodium Chloride

Topics: Adult; Antigens, Fungal; Aspergillosis; Galactose; HIV Infections; Humans; Male; Mannans; Middle Aged; Mycoses; Penicillium

We compared a real time-nucleic acid sequence-based amplification (RTi-NASBA) with conventional NASBA, galactomannan enzyme immunosorbent assay (GMEIA), and Mycology Study Group of the European Organization for Research and Treatment of Cancer (EORTC/MSG) criteria for the diagnosis of invasive aspergillosis (IA). From May 2004 to May 2005, blood samples (314 in total) were collected twice a week from 78 patients with hematologic diseases during neutropenic fever after chemotherapy or hematopoietic stem cell transplantation. Results were compared with each other on the basis of EORTC/ MSG criteria. The cutoff of conventional NASBA was set to be 3.5; GM 0.5; RTi-NASBA, 20% above the negative control. There were 22 patients with IA (7 probables and 15 possibles) and 56 patients with nonfungal infection. The Kappa statistic for RTi-NASBA versus conventional NASBA was 0.80 (0.66-0.82; p<0.001) indicating that there was fairly good accordance between two tests. RTi-NASBA showed sensitivity 0.96, specificity 0.43, positive- and negative-predictive value 0.40 and 0.96, respectively. GM showed good specificity (0.98), while the sensitivity (0.45) was poor. When we use the combination of GM with either of two NASBAs, the sensitivity was improved up to 100%. In conclusion, RTi-NASBA could be a good alternative to the conventional one for the screening of IA.

Topics: Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Nucleic Acid Amplification Techniques; Reproducibility of Results; RNA, Fungal; Sensitivity and Specificity

We assessed Calcofluor white staining, Aspergillus polymerase chain reaction, and a galactomannan enzyme immunoassay for diagnosis of fungal infection with use of computed tomography-guided percutaneous lung biopsy specimens obtained from 61 patients. The sensitivity and specificity of computerized tomography, Aspergillus polymerase chain reaction, and galactomannan enzyme immunoassay were 100% and 50%, 100% and 86%, and 88% and 94%, respectively.

Topics: Adult; Aspergillosis; Aspergillus; Benzenesulfonates; Biopsy; Contrast Media; False Positive Reactions; Female; Galactose; Humans; Immunocompromised Host; Immunoenzyme Techniques; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Prospective Studies; Sensitivity and Specificity; Tomography, X-Ray Computed

The diagnosis of invasive aspergillosis which is a serious infection of immunocompromized patients, depends on the detection of Aspergillus galactomannan antigen in the serum by enzyme immunoassay (EIA) in routine laboratories. However, it has been previously reported that false positive results in Aspergillus galactomannan test may be obtained in the sera of patients sera receiving piperacillin-tazobactam (PIP-TAZ). The aim of this study was to investigate the presence and levels of Aspergillus galactomannan antigen in the content of PIP-TAZ and some other antimicrobial agents that are often used for the treatment of infections in immunocompromised patients. The level of galactomannan antigen was determined for PIP-TAZ, ampicillin-sulbactam, ampicillin, penicillin G, ceftriaxone, cefepime, imipenem, clarithromycin, ciprofloxacin, vancomycin, gentamicin, trimethoprim-sulfamethoxazole, ornidazole, fluconazole and amphotericin B, by a commercial EIA (Platelia Aspergillus EIA, Bio-Rad, France) kit. Galactomannan index (GI) was estimated with the ratio of absorbance values of antimicrobials to cut-off value and evaluated as positive when GI was found >0.5. Amongst the 15 antibiotics studied, the only positive result was detected for ampicillin with the highest index value (GI = 0.540), followed by PIP-TAZ with a relatively high value (GI = 0.235) even though it was not in the range of positivity. GI values have ranged from 0.011 to 0.188 for the other antibiotics. In conclusion, the use of especially ampicillin (and probably PIP-TAZ) therapy should be questioned in patients whose sera are being tested for Aspergillus galactomannan antigen by EIA in order to evaluate the positive results in terms of false positivities due to cross reactivity.

Topics: Ampicillin; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Cross Reactions; Drug Contamination; False Positive Reactions; Galactose; Humans; Immunoenzyme Techniques; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination

To assess the value of Aspergillus galactomannan (GM) double-direct sandwich enzyme-linked immunosorbent assay (ELISA) in the diagnosis of invasive pulmonary aspergillosis (IPA).. Ninety adult SD rats were randomly divided into 5 groups, including 4 groups of pulmonary infection by Aspergillus fumigatus (A. fumigatus), Candida albicans, mucor, and Streptococcus pneumoniae, respectively, and a colonization group by A. fumigatus in the pharynx oralis (n = 18 each). For the infection models, suspensions of pathogenic bacteria and fungi were instilled into the lungs of the rats by tracheal intubation. For the colonization model, the suspension of A. fumigatus was applied to the nasal cavity and pharynx oralis of the rats. The animals were sacrificed on days 3, 7, and 12 after inoculation, and blood samples were obtained by cardiac puncture and used for GM detection. The lung tissues were prepared for routine pathology examination, and hexamethylene tetramine silver staining was used to detect the fungi. A double-direct sandwich ELISA was employed to detect GM optical density index in the serum samples.. The lung tissues of rats infected with A. fumigatus, Candida albicans, mucor and Streptococcus pneumoniae all showed remarkable inflammatory reactions, and hyphae were observed in rats with fungal infection (including A. fumigatus and mucor), and spores in rats infected with Candida albicans. The lung tissues of the A. fumigatus colonization rats showed no inflammatory reactions. The serum GM optical density index of the groups infected with A. fumigatus, Candida albicans, mucor and Streptococcus pneumoniae, and the A. fumigatus colonization group were 1.69 +/- 0.29, 0.89 +/- 0.46, 0.87 +/- 0.39, 0.77 +/- 0.34 and 0.90 +/- 0.49, respectively. The serum GM optical density index of the IPA group was higher than those of the other 4 groups (P < 0.05). If the cutoff ODI was 1.5, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for diagnosis of IPA in the rats were 78.6%, 87.5%, 57.9% and 94.9%, respectively. The sensitivity for A. fumigatus infection on days 3, 7, and 12 was 60.0%, 80% and 100%, and the specificity was 95.5%, 81.0% and 85.7%, respectively.. GM detection could distinguish Aspergillus infection from Candida albicans, mucor, Streptococcus pneumoniae infection and Aspergillus colonization. The sensitivity of the test for the diagnosis of IPA tended to be higher with longer duration of infection. A cutoff ODI of 1.5 showed the best sensitivity and specificity for IPA in this rat model.

Topics: Animals; Aspergillosis; Disease Models, Animal; Female; Galactose; Lung Diseases, Fungal; Male; Mannans; Rats; Rats, Sprague-Dawley; Sensitivity and Specificity

Topics: Antifungal Agents; Antigens, Fungal; Aspergillosis; Biomarkers; Clinical Trials as Topic; Galactose; Humans; Immunoglobulin E; Lung Diseases, Fungal; Lung Diseases, Interstitial; Mannans; Practice Guidelines as Topic; Pyrimidines; Serologic Tests; Triazoles; Voriconazole

Topics: Aspergillosis; Aspergillus; beta-Glucans; Blood; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Mycology; Polymerase Chain Reaction; Sensitivity and Specificity; Tomography, X-Ray Computed

Invasive aspergillosis (IA) is associated with high mortality. Successful outcome with treatment is linked to early diagnosis. The utility of classic diagnostic methods, however, is limited.. To aid in the diagnosis of IA, we retrospectively assessed our diagnostic service, using real-time polymerase chain reaction (PCR) and galactomannan sandwich enzyme-linked immunosorbent assay (ELISA).. A total of 203 patients at risk of invasive fungal infection were screened by PCR, and 116 of the patients were also tested by ELISA. The patient group comprised 176 patients with hematological malignancy and 28 control patients with evidence of invasive candidal infection. Consensus European Organisation for Research and Treatment of Cancer and Mycoses Study Group criteria were used to classify fungal infection, which, by definition, excluded the PCR result. The PCR method was sensitive (up to 92.3% sensitivity) and specific (up to 94.6% specificity) and had good agreement with the galactomannan ELISA (76.7%) and high-resolution computed tomography scan results.. A negative PCR result can be used to rule out IA and to limit the need for empirical antifungal therapy; thus, it has a role in diagnosing IA infections, especially in combination with antigen testing. PCR-positive cases classified as "false positives" regularly reflect the limitations of classic microbiological procedures or restricted use of consensus clinical methods employed to classify infection.

Topics: Adolescent; Adult; Antifungal Agents; Antineoplastic Agents; Aspergillosis; Aspergillus; Blood; Candidiasis; Child; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Hematologic Neoplasms; Humans; Mannans; Middle Aged; Opportunistic Infections; Polymerase Chain Reaction; Retrospective Studies; RNA, Fungal; Sensitivity and Specificity; Stem Cell Transplantation; Tomography, X-Ray Computed

Current laboratory and radiological methods for diagnosis of invasive aspergillosis (IA) lack sensitivity and specificity.. We prospectively evaluated the diagnostic value of twice-weekly screening for circulating Aspergillus fumigatus and A. flavus DNA with a polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA).. Among the 201 adult patients with hematological malignancies who were included in the study, 55 IA cases were diagnosed. On the basis of the analysis of 1205 serum samples from 167 patients, the sensitivity, specificity, and positive and negative predictive values of the PCR-ELISA for proven and probable IA cases were 63.6%, 89.7%, 63.6%, and 89.7%, respectively, when samples with 2 consecutive positive results were used. The use of a combination of the PCR-ELISA and a galactomannan (GM) assay increased the sensitivity to 83.3%, increased the negative predictive value to 97.6%, and decreased the specificity to 69.8%. In most patients with IA, PCR-ELISA positivity anticipated or was simultaneous with the initiation of antifungal therapy, the abnormalities found by computed tomography, the mycological/histological diagnosis, and the GM positivity. Overall, 56.3% of the patients had at least 1 positive sample, and the false single-positive rate was 44.8%.. In addition to serial screening for GM antigenemia and radiological surveillance, PCR-ELISA may improve the rates of early diagnosis of IA and the management of patients with hematological malignancies.

Topics: Aspergillosis; Aspergillus flavus; Aspergillus fumigatus; DNA, Fungal; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Galactose; Hematologic Neoplasms; Humans; Mannans; Molecular Diagnostic Techniques; Polymerase Chain Reaction; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity

Several reports have described a high rate of false-positive Aspergillus galactomannan (GM) test results for patients treated with piperacillin-tazobactam. In this retrospective study, we first examined the relationships between intravenous administration of three beta-lactam antibiotics and the occurrence of false-positive GM test results in hematology patients. We then estimated the kinetics of clearance of GM after the cessation of treatment. Sequential serum samples from 69 patients that had received beta-lactams were analyzed by using a Platelia Aspergillus test. A significant association was found between GM positivity (>/=0.5) and the administration of beta-lactams (P < 0.0001). The direct role of beta-lactams in patients' serum positivity was assessed by testing 39 batches of beta-lactams, of which 27 were positive for GM. None of the latter were positive according to a fungus- and Aspergillus-specific PCR. The kinetics of the decrease of GM was analyzed on sequential serum samples obtained after treatment. By use of a nonlinear regression model, the average time to negative antigen was assessed to be 5.5 days (95% confidence interval [CI], 4.1 to [7.0]), with a half-life of elimination of GM of 2.4 days (95% CI, 1.8 to 3.0). This study confirms that the administration of beta-lactams containing GM is responsible for false-positive diagnostic results, even up to 5 days after the cessation of treatment.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Aspergillus; beta-Lactams; Child; Child, Preschool; False Positive Reactions; Female; Galactose; Hematologic Diseases; Humans; Kinetics; Male; Mannans; Middle Aged; Treatment Outcome

A commercial sandwich elisa (Platelia Aspergillus EIA; Bio-Rad) developed for the detection of galactomannan, a major cell wall constituent of Aspergillus species, was tested for its efficacy in the diagnosis of aspergillosis in falcons. Ninety serum samples from 50 aspergillosis-positive falcons and 182 samples from 142 aspergillosis-negative falcons were tested. The sensitivity of the test was only 12 per cent and its specificity was 95 percent. The test was therefore unsatisfactory for detecting galactomannan in the serum samples and cannot be used as a screening test for aspergillosis in falcons.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus; Bird Diseases; Enzyme-Linked Immunosorbent Assay; Falconiformes; Galactose; Mannans; Sensitivity and Specificity

Topics: Academic Medical Centers; Aspergillosis; Aspergillus; Galactose; Hematologic Neoplasms; Immunoenzyme Techniques; Mannans; Opportunistic Infections

Topics: Aspergillosis; Galactose; Humans; Mannans; Reproducibility of Results

We present a case of primary hepatic and possible splenic invasive aspergillosis (IA), which progressed under anti-CD20 B cell treatment for posttransplantation lymphoproliferative disease following allogeneic stem cell transplantation, to highlight the clinical value of a positive galactomannan-antigen test, an intestinal portal of entry of Aspergillus, and the detrimental effect of B lymphocyte depletion in IA.

Topics: Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Antifungal Agents; Antineoplastic Agents; Aspergillosis; Chemical and Drug Induced Liver Injury; Fatal Outcome; Female; Galactose; Graft vs Host Disease; Humans; Liver Diseases; Lymphoproliferative Disorders; Mannans; Middle Aged; Pyrimidines; Rituximab; Splenic Diseases; Stem Cell Transplantation; Transplantation, Homologous; Triazoles; Voriconazole

Early diagnosis of invasive pulmonary aspergillosis (IPA) is important as prompt treatment with antifungal drugs may increase patient survival. Our study investigated the efficiency of routine testing of the Aspergillus galactomannan antigen (AGA) test in combination with chest CT scans for IPA diagnosis.. From February 2002 to June 2004, 74 hemato-oncologic patients undergoing allogeneic stem cell transplantation were prospectively studied with serum AGA twice weekly from admission until death or discharge and weekly afterward when possible. Chest CT scans were performed when fever of unknown origin had lasted beyond 3 days of antibacterial therapy.. Seven patients were classified with possible IPA and two patients, proven IPA. Fourteen patients showed positive results for AGA (OD index>or=1.0 on two subsequent sera). The sensitivity and specificity of the test were 100% and 93%, respectively; the positive and negative predictive values were 64% and 100%, respectively. All patients with possible/proven IPA showed abnormal CT signs; in four cases, imaging signs followed AGA positivity (median 5 days), whereas in five cases they preceded serologic positivity (median, 8 days). In the nine patients with IPA, antifungal therapy was promptly instituted, including lipid formulations of amphotericin B (n=5) or caspofungin (n=4). In only two of the nine patients (22%) with IPA, the primary cause of death was fungal infection.. The combination of AGA detection and early chest CT scans might be considered useful tools to detect minimal changes of IPA. Based on these findings, aggressive antifungal therapy should be initiated.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Galactose; Hematologic Neoplasms; Humans; Lung Diseases; Mannans; Middle Aged; Neoplasms; Retrospective Studies; Stem Cell Transplantation; Transplantation, Homologous; Treatment Outcome

Bronchoalveolar lavage (BAL) is widely used for evaluation of patients with suspected invasive pulmonary aspergillosis (IPA). However, the diagnostic yield of BAL for detection of IPA by culture and direct examination is limited. Earlier diagnosis may be facilitated by assays that can detect Aspergillus galactomannan antigen or DNA in BAL fluid. We therefore characterized and compared the diagnostic yields of a galactomannan enzyme immunoassay (GM EIA), quantitative real-time PCR (qPCR), and quantitative cultures in experiments using BAL fluid from neutropenic rabbits with experimentally induced IPA defined as microbiologically and histologically evident invasion. The qPCR assay targeted the rRNA gene complex of Aspergillus fumigatus. The GM EIA and qPCR assay were characterized by receiver operator curve analysis. With an optimal cutoff of 0.75, the GM EIA had a sensitivity and specificity of 100% in untreated controls. A decline in sensitivity (92%) was observed when antifungal therapy (AFT) was administered. The optimal cutoff for qPCR was a crossover of 36 cycles, with sensitivity and specificity of 80% and 100%, respectively. The sensitivity of qPCR also decreased with AFT to 50%. Quantitative culture of BAL had a sensitivity of 46% and a specificity of 100%. The sensitivity of quantitative culture decreased with AFT to 16%. The GM EIA and qPCR assay had greater sensitivity than culture in detection of A. fumigatus in BAL fluid in experimentally induced IPA (P+/-0.04). Use of the GM EIA and qPCR assay in conjunction with culture-based diagnostic methods applied to BAL fluid could facilitate accurate diagnosis and more-timely initiation of specific therapy.

Topics: Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Female; Galactose; Immunoenzyme Techniques; Lung Diseases, Fungal; Mannans; Polymerase Chain Reaction; Rabbits; Sensitivity and Specificity

Profound and prolonged neutropenia following chemotherapy is a major risk factor for systemic fungal infection. As the early diagnosis of invasive fungal infection (IFI) is difficult, these infections are still associated with high morbidity and mortality. Recently, Pan-fungal polymerase chain reaction (PCR) has been a promising aid in rapid, early diagnosis of IFI. During the past few years, increasing numbers of suspected IFIs were encountered at our institution in patients with prolonged neutropenia after intensified immunosuppressive chemotherapy. The aim of this study was to investigate the diagnostic utility of both the aspergillus galactomannan (GM) antigen and the pan-fungal PCR assay in the diagnosis of IFI in high risk febrile neutropenic paediatric cancer patients. During one year period, 91 febrile neutropenic (FN) paediatric cases at high risk for developing IFI while receiving chemotherapy were investigated at National Cancer Institute, Egypt. These patients were subjected to clinical evaluation, chest CT scan, conventional blood cultures for bacterial and fungal pathogens, aspergillus GM antigen detection and PCR assay utilizing pan-fungal primers. Of the 91 FN episodes, 15 were proven IFI; whereas 27 cases were either probable (n=13) or possible IFI (n=14), and 49 were unlikely to be IFI episodes. Based on positive results for proven/probable IFI and compared to culture results, Pan-fungal PCR showed sensitivity, specificity, positive and negative predictive values of 75%, 92%, 84% and 87%; respectively. Aspergillus antigen test showed a sensitivity of 79%, specificity of 61%, positive and negative predictive values of 54% and 83%; respectively. A negative PCR in the proven and probable cases was closely related to previous antifungal therapy for a prior history of IFI. In patients at high risk for IFI, neither the sensitivity, nor specificity of the GM test was sufficient. The results of PCR assay was reasonably specific but not very sensitive and had a chance of missing the diagnosis of IFI. The PCR assay seems a promising test for objectively defining IFI, but is not recommended as the only tool for diagnosing IFI. Combining microscopy, culture, and PCR may improve the diagnostic outcome.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Aspergillus; Blood; Child; Child, Preschool; Egypt; Electrophoresis, Agar Gel; Galactose; Humans; Mannans; Mycoses; Neoplasms; Polymerase Chain Reaction; Predictive Value of Tests; Sensitivity and Specificity; Statistics as Topic

Topics: Antigens, Fungal; Aspergillosis; Galactose; Humans; Mannans

Invasive aspergillosis (IA) is among the most common invasive fungal infections in neutropenic patients with hematological disorders in the authors' institution, King Chulalongkorn Memorial Hospital (KCMH), Bangkok, Thailand Previous studies have reported the Aspergillus galactomannan enzyme immunosorbent assay (GMEIA) may be a useful diagnostic tool for IA. The authors evaluated the performance of the GM EIA for the diagnosis and monitoring of the course of IA in KCMH.. The authors prospectively performed the study from June 2002 to January 2004 in a consecutive series of adult neutropenic patients with hematological disorders who were at risk for developing IA. During hospitalization, serum galactomannan levels were measured once or twice weekly using the Platellia Aspergillus EIA test kit. The sensitivity and specificity of the GM EIA were calculated according to the proportion of patients with true and false positive and negative tests.. There were 50 treatment episodes in 44 patients with 5 proven, 12 probable, and 33 possible or no IA. The cutoff GM index of > 0.75 was determined with a sensitivity of 94.1% and a specificity of 78.8%. There was a close relationship between clinical outcome and the kinetics of GM indices.. The GM EIA is a useful diagnostic toolfor the diagnosis and monitoring of the course oflA in the presented institute.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Diseases; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Neutropenia; Opportunistic Infections; Prospective Studies; Reagent Kits, Diagnostic; Risk; Sensitivity and Specificity

The incidence of invasive aspergillosis (IA) is increasing in patients with hematological disorders and it may lead to a high mortality rate. This study was to evaluate serum aspergillus galactomannan (GM) antigen assay as a potential early diagnosis and follow-up of IA.. From October 2004 to October 2005, 302 blood samples were obtained from 81 neutropenic hematological patients with fever over 38.5 degrees C and shown to have no response to broad-spectrum antibiotics treatment. Blood samples were collected twice a week. The detection of aspergillus GM antigen was carried out with a sandwich enzyme-linked immunosorbent assay and a GM positivity was defined as A index > 1.5 in two consecutive mensuration. Furthermore, the patients with positive GM test received preemptive antifungal therapy with amphotericin B or itraconazole.. Twenty seven patients (33.0%) were considered GM test positive from a total of 81 cases and 11 patients were diagnosed as proven or probable IA. The GM test correctly identified 7 of the 11 patients who had aspergillus antigen (63.6% sensitivity). When 14 patients without signs or symptoms of invasive fungal infection (IFI) were tested, the test correctly identified 12 of the 14 (85.7% specificity) as not having the antigen. GM positivity allowed also the anticipation of IA diagnosis (from 3 to 30 days before mycological culture). 19 of 27 GM test positive patients were given preemptive anti aspergillus therapy and there was a good response in 12 patients but no response in 7 cases with 36.8% mortality. After treatment, GM antigen decreased to normal with a good response. Otherwise, an elevated value hinted a unsatisfactory result as judged with Wilcoxon signed rank test (P < 0.0005).. It is suggested that serum GM antigen detection may be a useful test for early diagnosis of IA and GM test-based preemptive antifungal therapy in hematological patients at high risk can decrease mortality of IA substantively. Moreover, the dynamic change of GM test value can be useful for assessing therapeutic response.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Female; Galactose; Hematologic Diseases; Humans; Male; Mannans; Middle Aged

Two noninvasive diagnostic tests, (1-->3)-beta-D-glucan (BG) (Glucatell) and galactomannan (GM) (Platelia Aspergillus), were used retrospectively in a twice-weekly screening for the diagnosis of invasive aspergillosis (IA) in 40 treatment episodes (one hospital visit per patient) in 40 neutropenic adult patients at high risk for IA. Five proven IA cases, three probable IA cases, and three possible IA cases were diagnosed. Diagnostic levels of both BG and GM were detected in 100% of patients with proven IA cases and in 66% of patients with probable IA cases. The kinetics of both markers in patients with IA were similar. The sensitivity, specificity, and positive and negative predictive values for GM and BG were identical, namely, 87.5, 89.6, 70, and 96.3%, respectively. False-positive reactions occurred at a rate of 10.3% in both tests, but the patients showing false-positive results were different in each test. Both tests anticipated the clinical diagnosis, computed tomography abnormalities, and the initiation of antifungal therapy in most patients, but BG tended to become positive earlier than GM. A combination of the two tests improved the specificity (to 100%) and positive predictive value (to 100%) of each individual test without affecting the sensitivity and negative predictive values. In conclusion, BG and GM detection are useful tests for the diagnosis of IA in high-risk hematological patients, but a combination of the two tests was very useful to identify false-positive reactions by each test.

Topics: Adolescent; Adult; Aged; Antifungal Agents; Aspergillosis; Aspergillus; beta-Glucans; Chromogenic Compounds; Female; Galactose; Humans; Male; Mannans; Middle Aged; Neutropenia; Predictive Value of Tests; Sensitivity and Specificity

Topics: Adult; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Gastrointestinal Diseases; Graft vs Host Disease; Humans; Mannans; Reproducibility of Results; Soybean Proteins

We evaluated nucleic acid sequence-based amplification (NASBA) and a galactomannan enzyme immunosorbent assay (GM-EIA) for the diagnosis of invasive aspergillosis (IA) in neutropenic febrile patients and for monitoring of its clinical course and outcome. Blood samples were collected twice per week from 128 patients with hematologic diseases during periods of neutropenic fever after undergoing chemotherapy or hematopoietic stem cell transplantation. A total of 448 blood samples were tested.. There were 14 patients with IA (2 patients with proven IA and 12 with probable IA). The median index of the initial NASBA in the IA group was more than 10-fold higher than that in the non-IA group. Galactomannan antigenemia (index, >0.5) was detected with a sensitivity of 86%. In receiver-operator characteristic analysis, the cutoff index of NASBA for the presumptive diagnosis of IA was determined to be 5.0. Combination of these 2 parameters (either a GM-EIA index of >or=0.5 or a NASBA index of >or=5.0) improved the sensitivity of diagnosis to 100%. There was a close relationship between patient outcome and the kinetics of NASBA values: failure of negative conversion during treatment resulted in death in almost all cases.. If either GM-EIA or NASBA results suggest IA, the diagnostic yield for IA could be improved, and NASBA could be a useful marker for predicting the clinical course and outcome of treatment.

Topics: Adult; Aged; Antifungal Agents; Aspergillosis; Female; Galactose; Hematologic Diseases; Humans; Immunoenzyme Techniques; Leukemia, Myeloid, Acute; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Myelodysplastic Syndromes; Nucleic Acid Amplification Techniques; Precursor Cell Lymphoblastic Leukemia-Lymphoma; Sensitivity and Specificity

Topics: Adolescent; Adult; Aged; Aspergillosis; Candidiasis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Middle Aged; Prospective Studies; Sensitivity and Specificity; Single-Blind Method

There has been minimal clinical experience with the use of the Aspergillus galactomannan enzyme-linked immunosorbent assay (ELISA) for patients receiving echinocandin therapy. We reviewed the experience with the galactomannan ELISA for 17 patients in a study of caspofungin treatment for invasive aspergillosis. The rate of successful outcomes for these patients was similar to that overall for participants in the study. Trends in antigenemia levels correlated with clinical and radiographic findings.

Topics: Adult; Aged; Antifungal Agents; Antigens, Fungal; Aspergillosis; Caspofungin; Echinocandins; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Lipopeptides; Lung Diseases, Fungal; Mannans; Middle Aged; Peptides, Cyclic; Treatment Outcome

A case of autopsy-proven fungal pneumonia in a relapsed leukaemia patient is reported. The fungus Hormographiella aspergillata was cultured from two bronchoalveolar fluid samples and identified through morphological examination and ITS2 sequence analysis. In addition, galactomannan was detected in eight consecutive serum samples, which suggested a co-infection with Aspergillus species. The patient was treated with caspofungin.

Topics: Adult; Agaricales; Aspergillosis; Bronchoalveolar Lavage Fluid; Coprinus; DNA, Intergenic; Fatal Outcome; Galactose; Humans; Leukemia, Myelomonocytic, Acute; Lung; Lung Diseases, Fungal; Male; Mannans; Tomography, X-Ray Computed

Until recently, accurate microbiological diagnosis of invasive aspergillosis (IA) was seldom established in HSCT recipients. Blood samples are rarely positive for Aspergillus species, the reliability of the cultures depends of the specimen (if taken from a normally sterile site or not) and biopsy samples require invasive procedures, rarely recommended in patients with severe thrombocytopenia. Implementing the international consensus defining the microbiological criteria for the diagnosis of Aspergillus infection, we retrospectively evaluated the role of serum galactomannan (GM) detection by EIA to diagnose IA among HSCT patients with proven invasive fungal infection (IFI) and the impact of serum storage in GM concentrations. The EIA assay allowed categorizing as "probable" 5 of the 10 cases of "possible" aspergillosis (50%). Considering a lower cut-off level for the reaction (1.0), 80% of the cases could be categorized as "probable" aspergillosis. Positive or undetermined results were detected one to 4 months before the diagnosis of IA in eight of the 11 patients (72.7%) with proven IFI. Retesting the stored samples after a second storage for four years, we could observe lower reactivity in 20% of the samples. The detection of galactomannan by the EIA test represents a major advance in the diagnosis of IA in HSCT recipients at high risk of IA. A better understanding of the kinetics of the GM in different clinical situations is necessary to maximize the benefit of the test in Aspergillus surveillance.

Topics: Adolescent; Adult; Aspergillosis; Aspergillus; Child; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunoenzyme Techniques; Male; Mannans; Retrospective Studies; Specimen Handling

We describe the case of a patient with improving invasive aspergillosis and paradoxically rising serum galactomannan levels in the presence of chronic renal failure and ongoing hemodialysis. Dialysate tested negative for galactomannan, demonstrating the inability of treatments such as hemodialysis to clear Aspergillus antigen from serum. In patients with renal failure and aspergillosis, rising serum galactomannan levels may not necessarily signify progressive infection.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Aspergillus flavus; Chronic Disease; Fungemia; Galactose; Humans; Male; Mannans; Renal Dialysis; Renal Insufficiency

In view of the poor therapy outcomes of invasive aspergillosis, the objective of this study was to evaluate the efficacy of combination treatment consisting of the polyene amphotericin-B-intralipid, the echinocandin caspofungin and granulocyte-colony stimulating factor (G-CSF) in experimental murine systemic aspergillosis. With inhibition of synthesis of 1,3-beta-d-glucan in the fungal cell wall by caspofungin and an effect on the cell membrane by amphotericin-B-intralipid, this treatment may result in a synergic effect against Aspergillus fumigatus. Addition of G-CSF may further contribute to therapy of aspergillosis.. ICR mice were immunosuppressed by intraperitoneal administration of cyclophosphamide. Three days later, the mice were inoculated intravenously (iv) with A. fumigatus conidia. Infection and treatment were evaluated during an observation period of 30 days in terms of mortality (survival rate and mean survival time) and morbidity (quantitative determination of fungal burden, histopathology, and detection of serum galactomannan).. Combination of caspofungin + G-CSF or addition of G-CSF to the combination of caspofungin + amphotericin-B-intralipid increased the survival rate of infected mice up to 78.9% and prolonged their mean survival time to 25 days. These combinations also resulted in a reduction in fungal burden in organs, and a decrease in serum galactomannan.. The successful results obtained in the experimental model may possibly open the way to more effective management of aspergillosis in humans.

Topics: Amphotericin B; Animals; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Caspofungin; Disease Models, Animal; Drug Synergism; Drug Therapy, Combination; Echinocandins; Female; Galactose; Granulocyte Colony-Stimulating Factor; Immunocompromised Host; Kidney; Lipopeptides; Liposomes; Lung; Mannans; Mice; Mice, Inbred ICR; Peptides, Cyclic; Polyenes; Survival Rate

Although invasive aspergillosis is recognized as an increased concern for immunosuppressed patients, establishing an early diagnosis remains a challenge. Current methods for diagnosis, including sensitive radiography and invasive procedures to identify organisms in affected tissues, provide an early diagnosis for only a small proportion of patients infected with Aspergillus species. The availability of a standardized assay to detect circulating Aspergillus galactomannan, the BioRad Platelia Aspergillus galactomannan enzyme immunoassay, should be considered as a valuable tool. However, at present, clinical utility is hampered by controversies surrounding the performance of the test and an apparent lack of definitive data. These controversies emphasize the complexity inherent in analyzing diagnostic tests for aspergillosis. The widespread availability of a standardized assay may now allow us to address the multiple sources of variability and bias via large, cooperative studies.

Topics: Aspergillosis; Aspergillus; Clinical Laboratory Techniques; Evaluation Studies as Topic; Galactose; Humans; Immunoenzyme Techniques; Mannans

Topics: Anti-Bacterial Agents; Aspergillosis; Drug Contamination; Galactose; Humans; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination

The Platelia galactomannan enzyme immunoassay is a commercially available nonculture method for diagnosing invasive aspergillosis. Recently, steps have been taken to improve sensitivity; specifically, a low (0.5 to 0.7) galactomannan index (GMI) value to determine positivity has been validated by multiple groups. We evaluated the intra-assay and interassay reproducibility at low index levels using three different kit lots on three different days in three different microbiology laboratories. Clinical and spiked sera were blinded and sent with galactomannan enzyme immunoassay (EIA) kits to the participating laboratories. We also prospectively collected data on all galactomannan EIAs performed between 1 September 2003 and 21 July 2004 at the University of Washington Medical Center microbiology laboratory to assess reproducibility of clinical samples analyzed in "real time." From the multilaboratory study, a total of 836 results were available for evaluation. Reproducibility was excellent between laboratories and on different days. Significant variability was seen between runs/lots, which may in part be associated with different threshold control values in different kits. Among the 1,410 clinical samples that were prospectively analyzed, 168 (90%) were confirmed to be positive on repeat testing (GMI, > or =0.5). Among the 19 (10.2%) initially positive samples not confirmed on repeat testing, the majority had a GMI at the threshold of the assay (between 0.5 and 0.7). Our findings suggest that the Platelia galactomannan immunoassay has good reproducibility. However, changes in GMI levels when different kit lots are used, and single samples with low-positive (GMI of 0.5 to 0.7) indices, should be interpreted with caution.

Topics: Antigens, Fungal; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Immunoenzyme Techniques; Laboratories; Mannans; Reagent Kits, Diagnostic; Reproducibility of Results; Sensitivity and Specificity

Detection of serum galactomannan (GM) antigen and presence of the halo sign on a pulmonary computerized tomographic (CT) scan have a high specificity but a low sensitivity to diagnose invasive aspergillosis (IA) in patients at risk for this disease. To our knowledge, the relationship between the time at which pulmonary infiltrates are detected by CT and the time at which GM antigens are detected by enzyme immunoassay (EIA) has not been studied.. In a prospective study, tests for detection of GM were performed twice weekly for patients with hematological malignancies who had undergone hematopoetic stem cell transplantation (HSCT) or had received induction and/or consolidation chemotherapy. A pulmonary CT scan was performed once weekly. Infiltrates were defined as either major or minor signs. IA was classified as proven, probable, or possible, in accordance with the definition stated by the European Organization for Research and Treatment of Cancer-Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group.. We analyzed 161 episodes of infection in 107 patients (65 allogeneic HSCT recipients, 30 autologous HSCT recipients, and 66 induction and/or consolidation chemotherapy recipients). A total of 109 episodes with no IA, 32 episodes with possible IA, and 20 episodes with probable or proven IA were identified. Minor pulmonary signs were detected by CT in 70 episodes (43%), and major pulmonary signs were detected by CT in 11 episodes (7%). Univariate and multivariate analyses revealed no significant association between detection of GM by EIA and detection of abnormal pulmonary signs by CT. A significant association was found between GM levels and receipt of piperacillin-tazobactam. GM test results were not positive before major signs were seen on CT images. Only 7 (10%) of 70 patients with minor pulmonary signs had positive GM test results before detection of the greatest pathologic change by CT.. We show that detection of GM by EIA does not precede detection of major lesions by pulmonary CT. In the clinical setting, the decision to administer mold-active treatment should based on detection of new pulmonary infiltrates on CT performed early during infection, rather than on results of EIA for detection of GM.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Aspergillosis; Female; Galactose; Hematologic Neoplasms; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Multivariate Analysis; Odds Ratio; Sensitivity and Specificity; Tomography, X-Ray Computed

To compare PCR with galactomannan antigen detection for the diagnosis of invasive aspergillosis (IA).. We prospectively collected serial blood samples from haematological patients at risk of IA, and analysed their samples retrospectively for galactomannan (GM) antigen using the Platelia test and for aspergillus DNA using an in-house PCR-ELISA assay. Matched GM and PCR analyses were performed on 263 samples from 25 patients. Patients were classified for potential IA according to international consensus criteria, with five patients classified as positive (four proven, one probable) and 20 classified as negative (seven possible, 13 no evidence IA).. All five patients with IA were positive by PCR with positive results in 24 of 82 samples, whereas three of five patients were positive by GM with four of 82 samples being positive. Three of 20 patients without IA were positive by PCR in 18 of 181 samples, whereas corresponding results for GM detection were one of 20 and one of 181, respectively. Adjustment of ELISA cut-off values and/or the requirement for two consecutive samples to be positive generated different results; however, lowering the positivity index (PI) for GM detection to 0.5 did not improve the sensitivity of the assay. Optimal results for PCR detection and GM were: 100% and 60% sensitivity, 85% and 95% specificity, 0.625 and 0.75 positive predictive value, and 1.0 and 0.8 negative predictive value, with a false-positive sample rate of 8 and 0.4%, positive likelihood ratio of 6.66 and 11.99 and negative likelihood ratio of 0 and 0.42, respectively.. This PCR method is very sensitive for the diagnosis of IA but is associated with a moderate rate of false positives; the GM assay exhibited poor sensitivity but high specificity. Further evaluation of PCR assays for the diagnosis of IA and other invasive fungal infections is warranted.

Topics: Adolescent; Adult; Aged; Animals; Antigens, Fungal; Aspergillosis; Aspergillus; Child, Preschool; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Reproducibility of Results; Sensitivity and Specificity

Empirical antifungal therapy is the standard treatment for persistent or relapsing antibiotic-resistant neutropenic fever. However, overtreatment resulting in increased toxicity and treatment-related cost is a major shortcoming of such therapy. We assessed the feasibility of a "preemptive" approach based on the incorporation of sensitive, noninvasive diagnostic tests for consecutive high-risk neutropenic patients who had received fluconazole prophylaxis while avoiding empirical therapy.. A total of 136 treatment episodes for persons who were at risk of acquiring invasive fungal infection (IFI) were screened for the presence of galactomannan with an enzyme immunoassay. A diagnostic evaluation, which included thoracic computed tomography scanning (HRCT) and bronchoscopy with lavage, was performed on the basis of well-defined clinical, radiological, and microbiological criteria. Only seropositive patients and patients with a positive microbiological test result plus supportive radiological findings received liposomal amphotericin B.. Neutropenic fever developed in 117 episodes, of which at least 41 episodes (35%) satisfied existing criteria for empirical antifungal therapy. However, our protocol-driven preemptive approach reduced the rate of antifungal use for these episodes from 35% to 7.7% (a 78% reduction) and led to the early initiation of antifungal therapy in 10 episodes (7.3%) that were clinically not suspected of being IFI. No undetected cases of invasive aspergillosis were identified; 1 case of zygomycosis was missed. Breakthrough candidemia was diagnosed by conventional culture techniques and was treated successfully. With use of a preemptive approach, the 12-week survival rate for patients with IFI was 63.6% (it was 63.1% for those with invasive aspergillosis).. Preemptive therapy based on enzyme immunoassay and HRCT reduced the exposure to expensive and potentially toxic drugs and offered effective antifungal control, but it failed to detect non-Aspergillus IFI.

Topics: Adolescent; Adult; Aged; Algorithms; Amphotericin B; Antifungal Agents; Aspergillosis; Feasibility Studies; Female; Galactose; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Mycoses; Neutropenia; Prospective Studies; Risk Factors; Tomography, X-Ray Computed

Positive galactomannan (GM) anti-genemias are included as a microbiological item in the diagnosis of probable or possible invasive aspergillosis (IA). Because false-positive GM results frequently occur, at least two positive results on two different samples are required. Waiting for clinical specimens can delay the initiation of treatment. As an alternative, we wondered whether detection of circulating Aspergillus DNA on the first positive GM serum sample could aid in diagnosing IA. Therefore, we retrospectively screened the first GM-positive serum samples from 29 patients from our hematology unit for Aspergillus DNA using real-time PCR. We compared the real-time PCR results with the final classification of proven, probable, and possible IA according to consensual criteria. No clear correlation between PCR results and the classification with the medical files could be shown. However, a positive PCR result was associated with a poor prognosis (Fisher's test; P=0.01). Our preliminary data suggest that a positive PCR result could indicate a more advanced stage of the disease. Therefore, concomitant positive PCR and GM results may justify the initiation of antifungal therapy in neutropenic patients. In contrast, a negative PCR on the first positive GM sample may argue for postponing costly antifungal administration until additional arguments for the diagnosis of IA are presented.

Topics: Adolescent; Adult; Aged; Antifungal Agents; Antigens, Fungal; Aspergillosis; Child; Child, Preschool; DNA, Fungal; Fungemia; Galactose; Humans; Mannans; Middle Aged; Polymerase Chain Reaction; Prognosis; Retrospective Studies

Detection of Aspergillus galactomannan (GM) in serum with the Platelia Aspergillus enzyme immunoassay (EIA) is useful for diagnosing invasive aspergillosis. From May 2003 to November 2004, 65 patients who did not develop aspergillosis had at least two positive sera while receiving a beta-lactam treatment (GM index [GMI], >or=0.5). Of the 69 treatment episodes scored, 41 consisted of a beta-lactam other than piperacillin-tazobactam (n=29), namely, amoxicillin-clavulanate (n=25), amoxicillin (n=10), ampicillin (n=3), or phenoxymethylpenicillin (n=2). In all cases, antigenemia became negative 24 h to 120 h upon stopping the antibiotic. Monitoring of 35 patients, including 26 with hematological malignancies, revealed three antigenemia kinetic patterns: each was observed with any drug regimen and consisted of a persistent GMI of >2.0 (65.7%), >0.5, and

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Lactams; False Positive Reactions; Fungemia; Galactose; Humans; Mannans

The performance of different in vitro diagnostic tests for the diagnosis of invasive aspergillosis (IA) was investigated in a transiently neutropenic rat model. Rats were immunosuppressed with cyclophosphamide and then inoculated intravenously with 1.5 x 10(4) CFU Aspergillus fumigatus spores. Animals were then either treated with caspofungin acetate, 1 mg/kg/day for 7 days, or not treated. PCR-enzyme-linked immunosorbent assay (ELISA), real-time PCR, and galactomannan (GM) detection were performed on postmortem blood samples, along with culture of liver, lung, and kidney homogenate. Caspofungin-treated animals showed a decrease in residual tissue burden of A. fumigatus from organ homogenate compared to untreated animals (P < 0.002). PCR-ELISA returned positive results for 11/17 animals treated with antifungal agents and for 10/17 untreated animals. Galactomannan was positive in 8/17 caspofungin-treated animals and 4/17 untreated animals. Real-time PCR was positive in 2/17 treated and 3/17 untreated animals. This study demonstrates that PCR-ELISA is a more sensitive test than either GM detection (P = 0.052) or real-time PCR (P < 0.01) for diagnosis of IA but that any of the three tests may return false-negative results in cases of histologically proven disease. Galactomannan indices from animals treated with antifungal agents showed a trend (P = 0.1) towards higher levels than those of untreated animals, but no effect was observed with PCR-ELISA indices (P = 0.29). GM detection, as previously described, may be enhanced by the administration of caspofungin, but PCR-ELISA appears not to be affected in the same way. We conclude that PCR-ELISA is a more sensitive and reliable method for laboratory diagnosis of IA.

Topics: Animals; Antifungal Agents; Aspergillosis; Caspofungin; Disease Models, Animal; Echinocandins; Enzyme-Linked Immunosorbent Assay; Galactose; Lipopeptides; Liver Diseases; Male; Mannans; Neutropenia; Peptides, Cyclic; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction

The utility of galactomannan antigen for diagnosing invasive aspergillosis was evaluated in 154 liver transplant recipients. Sample agreement was 98.5%, and patient specificity was 87%. Galactomannan positivity correlated with mortality, even when controlled for the number of tests performed. Whether galactomannan positivity identifies a subgroup at risk for poor outcome warrants further evaluation.

Topics: Adult; Aged; Aspergillosis; Female; Galactose; Humans; Immunoenzyme Techniques; Liver Transplantation; Male; Mannans; Middle Aged

Three techniques for the diagnosis of mammary aspergillosis in ewes were compared: indirect ELISA to detect the level of anti-Aspergillus IgG in serum, determination of galactomannan (Platelia procedure), and detection of DNA of Aspergillus in serum by a nested PCR. Twenty sera from proven cases of aspergillosis in ewes were positive using ELISA (100%), 80% were positive using PCR, but only 55% were positive using Platelia. All 20 control sera were negative using ELISA and PCR, whereas using Platelia methodology one was positive and the other doubtful. The detection of antibody by ELISA in sera is therefore a reliable criterion for the diagnosis of mammary aspergillosis in ewes. Platelia showed the same deficiencies reported in humans, with the appearance of false positives and negatives. The use of PCR was promising and might have valuable application in human medicine.

Topics: Animals; Antibodies, Fungal; Aspergillosis; Aspergillus fumigatus; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Mannans; Mastitis; Polymerase Chain Reaction; Sheep; Sheep Diseases

A major difficulty with the detection of circulating galactomannan, a cell-wall polysaccharide released by Aspergillus sp during growth, in the serodiagnosis of invasive aspergillosis is the occurrence of false-positive ELISA results, especially in neonates and infants. On the basis of molecule similarity, we postulate that a lipoteichoic acid of Bifidobacterium sp can act as epitope for the monoclonal antibody used in the ELISA. The neonatal gut is heavily colonised with Bifidobacterium sp and these bacteria or their lipoteichoic acid might cause ELISA reactivity with serum after translocation because of immaturity of the intestinal mucosa. If our hypothesis is correct, we might find a method to discriminate between false-positive and true-positive ELISA results and thereby prevent unnecessary pre-emptive treatment of patients.

Topics: Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus; Bifidobacterium; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Epitopes; False Negative Reactions; False Positive Reactions; Galactose; Humans; Infant; Infant, Newborn; Intestinal Mucosa; Lipopolysaccharides; Mannans; Models, Immunological; Serologic Tests; Teichoic Acids

Invasive aspergillosis (IA) is a considerable clinical problem in neutropenic patients with haematological malignancies but its diagnosis remains difficult. We prospectively evaluated a LightCycler polymerase chain reaction (PCR) assay, a nested-PCR assay and a galactomannan (GM) enzyme-linked immunosorbent assay (ELISA) to validate their significance in diagnosing IA. During 205 treatment episodes in 165 patients from six centres, a nested-PCR assay and GM testing was performed at regular intervals. Positive nested-PCR results were quantified by a LightCycler PCR assay. Patient episodes were stratified according to the 2002 European Organization for Research and Treatment of Cancer/Mycosis Study Group consensus criteria and the PCR and serology results were correlated with the clinical diagnostic classification. Sensitivity and specificity rates for the nested-PCR assay were up to 63.6% [95% confidence interval (CI): 30.8-89%) and 63.5% (95% CI: 53.4-72.7%) respectively, and 33.3% and 98.9% (95% CI: 7.5-70.1% and 94.2-99.9%) for GM respectively. The LightCycler PCR assay yielded positive results in 21.4%, lacking discrimination by quantification across the different clinical categories. In this prospective comparison, PCR was superior to GM with respect to sensitivity rates. In patients at high risk for IA, positive results for Aspergillus by PCR of blood samples are highly suggestive for IA and contribute to the diagnosis.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Neutropenia; Polymerase Chain Reaction; Prospective Studies

The clinical utility of Platelia trade mark Aspergillus galactomannan antigen for the early diagnosis of invasive aspergillosis was prospectively assessed in 70 consecutive lung transplant recipients. Sera were collected twice weekly and tested for galactomannan. Invasive aspergillosis was documented in 17.1% (12/70) of the patients. Using the generalized estimating equation model, at the cutoff value of >or= 0.5, the sensitivity of the test was 30%, specificity 93% with positive and negative likelihood ratios of 4.2 and 0.75, respectively. Increasing the cutoff value to >or= 0.66 yielded a sensitivity of 30%, specificity of 95%, and positive and negative likelihood ratios of 5.5 and 0.74. A total of 14 patients had false-positive tests, including nine who had cystic fibrosis or chronic obstructive pulmonary disease. False-positive tests occurred within 3 days of transplantation in 43% (6/14) of the patients, and within 7 days in 64% (9/14). Thus, the test demonstrated excellent specificity, but a low sensitivity for the diagnosis of aspergillosis in this patient population. Patients with cystic fibrosis or chronic obstructive pulmonary disease may transiently have a positive test in the early post-transplant period.

Topics: Adult; Aged; Antigens; Aspergillosis; Aspergillus; False Positive Reactions; Female; Galactose; Humans; Immunoenzyme Techniques; Lung; Lung Transplantation; Male; Mannans; Middle Aged

Invasive aspergillosis (IA) has become the leading infectious cause of death after allogeneic hematopoietic stem cell transplantation (allo-HSCT). This is partially because of the lack of a sensitive, specific, and noninvasive diagnostic test. New diagnostic tests for IA, such as the detection of Aspergillus galactomannan antigen (AGA) by sandwich enzyme-linked immunoabsorbent assay (ELISA), have recently been described. This study validates the usefulness of this diagnostic tool in the allo-HSCT setting.. From January 1999 to January 2001, all consecutive adult patients undergoing allo-HSCT were prospectively studied with a galactomannan antigenemia assay (ELISA test) twice weekly from admission until death or discharge, and weekly afterward if the patient received immunosuppressive therapy. Proven, probable, and possible IA were defined according to the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria.. During the 2 years of study, 74 patients underwent an allo-HSCT. A total of 832 serum samples were collected. According to the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria, it was ascertained that 66 patients did not fulfill any criteria of IA, 2 patients were classified with possible IA, 5 patients were classified with probable IA, and 1 patient was classified with proven IA. Fourteen samples were positive for AGA, all from patients with IA. The sensitivity and specificity of the test were 75% and 100%, respectively. The positive predictive and negative predictive values were 100% and 97%, respectively.. In this study, AGA detection was clearly related to IA. Although the ELISA test did not have any role in the anticipation of the diagnosis, it clarifies the diagnosis of IA in allo-HSCT.

Topics: Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunocompromised Host; Mannans; Predictive Value of Tests; Prospective Studies; Transplantation, Homologous

This report describes the use of the polymerase chain reaction (PCR) and galactomannan detection to detect aspergillus in the continuous ambulatory peritoneal dialysis (CAPD) fluid and blood of a patient with multiple myeloma on CAPD and immunosuppressive treatment. Diagnosis of aspergillosis was initially made by conventional culture of CAPD fluid, but the PCR and galactomannan assays also detected aspergillus DNA and antigen in the blood, respectively. This suggests that the PCR and galactomannan assays, previously suggested as useful in the management of invasive fungal infections in neutropenic haematological patients, may be suitable for application to a broad range of clinical situations and sample types.

Topics: Aspergillosis; Aspergillus fumigatus; Galactose; Humans; Male; Mannans; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritonitis; Polymerase Chain Reaction

The establishment of an optimal noninvasive method for diagnosing invasive aspergillosis (IA) is needed to improve the management of this life-threatening infection in patients with hematological disorders, and a number of noninvasive tests for IA that target different fungal components, including galactomannan, (1-->3)-beta-d-glucan (BDG), and Aspergillus DNA, have been developed. In this study, we prospectively evaluated the diagnostic potential of three noninvasive tests for IA that were used in a weekly screening strategy: the double-sandwich enzyme-linked immunosorbent assay (ELISA) for galactomannan (Platelia Aspergillus), a real-time PCR assay for Aspergillus DNA (GeniQ-Asper), and an assay for BDG (beta-glucan Wako). We analyzed 149 consecutive treatment episodes in 96 patients with hematological disorders who were at high risk for IA and diagnosed 9 proven IA cases, 2 probable IA cases, and 13 possible invasive fugal infections. In a receiver-operating characteristic (ROC) analysis, the area under the ROC curve was greatest for ELISA, using two consecutive positive results (0.97; P = 0.036 for ELISA versus PCR, P = 0.055 for ELISA versus BDG). Based on the ROC curve, the cutoff for the ELISA could be reduced to an optical density index (O.D.I.) of 0.6. With the use of this cutoff for ELISA and cutoffs for PCR and BDG that give a comparable level of specificity, the sensitivity/specificity/positive predictive value/negative predictive value of the ELISA and the PCR and BDG tests were 1.00/0.93/0.55/1.00, 0.55/0.93/0.40/0.96, and 0.55/0.93/0.40/0.96, respectively. In conclusion, among these weekly screening tests for IA, the double-sandwich ELISA test was the most sensitive at predicting the diagnosis of IA in high-risk patients with hematological disorders, using a reduced cutoff of 0.6 O.D.I.

Topics: Adolescent; Adult; Aged; Aspergillosis; beta-Glucans; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Glucans; Hematologic Diseases; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Prospective Studies; ROC Curve; Sensitivity and Specificity

Invasive aspergillosis (IA) is a frequent complication of blood or marrow transplantation. Previous studies have reported that the Aspergillus galactomannan enzyme immunoassay (GM EIA) may be a useful diagnostic tool for IA, but its sensitivity is variable. We examined the performance of the GM EIA in 986 serum samples from 67 patients. Results demonstrated that decreasing the index cutoff for positivity to 0.5 increased its sensitivity, with minimal loss of specificity. The low cutoff increased the duration of test positivity before diagnosis by clinical means. Sensitivity was highest in patients who did not receive preventative mold-active antifungals (87.5%). A rabbit model demonstrated that the level of circulating antigen correlated with the tissue fungus burden. A quantifiable response to antifungal therapy in clinical samples and the rabbit model supports the development of this assay for early diagnosis and therapeutic monitoring. The 0.5 cutoff may allow for better performance as an early diagnostic test.

Topics: Adolescent; Adult; Aged; Animals; Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Child; Child, Preschool; Disease Models, Animal; Female; Fungemia; Galactose; Humans; Immunoenzyme Techniques; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Rabbits; Sensitivity and Specificity

The recent advent of an improved commercial serum enzyme-linked immunosorbent assay (ELISA) for the detection of circulating galactomannan (GM), a major constituent of Aspergillus cell walls, has contributed to the diagnosis of invasive aspergillosis (IA) in many haematology and transplant centres. However, the optimal threshold for positivity remains a matter of debate. We prospectively evaluated the impact of lowering the cut-off in 124 neutropenic episodes with a high pretest probability for IA. Two new cut-off points, lower than previously accepted, are proposed: (a) a 'static' cut-off at 0.8 and (b) a 'dynamic' cut-off at 0.5. A single assay with an optical density (OD) index > or = 0.8 warrants the initiation of anti-Aspergillus therapy. A further lowering of the 'static' threshold seems not clinically feasible given the drop in positive predictive value (PPV). However, the demonstration of at least two sequential sera with an OD > or = 0.5 ('dynamic' threshold) increased the specificity and the PPV to 98.6% and the efficiency to 98%. Applying both cut-offs to a subgroup of 21 'possible' fungal infections further identified and upgraded six cases of IA. However, the clinical benefit of lower cut-offs (particularly for earlier diagnosis) depends upon the kinetics of antigenaemia and the intensity of serum sampling.

Topics: Adolescent; Adult; Aged; Aspergillosis; Biomarkers; Enzyme-Linked Immunosorbent Assay; Feasibility Studies; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Neutropenia; Opportunistic Infections; Prospective Studies; Sensitivity and Specificity

We analyzed the performance of (1, 3)-Beta-D-glucan (measurement by the alkaline-kinetic chromogenic Limulus method (FUNGITEC G test-MK, Fungitec) and the kinetic turbidimetric Limulus method [Beta-Glucan test WAKO, Wako]) and we carried out Aspergillus galactomannan antigen detection (enzyme-linked immunosorbent assay, ELISA test) for the early diagnosis of invasive aspergillosis in patients with hematological diseases at the time of febrile episodes of unknown origin that did not respond to antibacterial therapy for more than 3 days. During a one-year period (April 2002 to March 2003), a total of 69 febrile episodes in 58 patients were studied; 8 cases of invasive aspergillosis were diagnosed according to the definition of the European Organization for Research and Treatment of Cancer/Mycosis Study Group, and 61 cases were found to be non-mycotic diseases. Based on the analysis of 69 results with confirmed disease status, the overall performance of the Fungitec, the Wako, and the ELISA test were as follows: sensitivity was 0.88, 0.63, and 0.50, respectively, whereas the specificity was 0.85, 0.98, and 1.0, respectively. Moreover, there was a strong relationship between the log-transformed values of the (1, 3)-Beta-D-glucan levels measured by the two methods (r = 0.92 [95%CI, 0.89-0.94] ; p<0.001). For the statistical analysis of these serological tests a receiver operating characteristic curve (ROC) was used, as well as the resulting area under the ROC curve (ROC AUC). The ROC AUC and the cut-off values that gave the highest accuracy were as follows: 0.92, 24.9 pg/ml for the Fungitec, 0.84, 7.3 pg/ml for the Wako, and 0.89, 0.9 COI for the ELISA test, respectively. In conclusion, these results indicate that both of the two (1, 3)-Beta-D-glucan measurement approaches served equally well as surveillance tools for determining the extent of invasive aspergillosis; in addition, the log-transformed value of these tests can be used for comparison. Moreover, the ELISA test was found to have clinical utility, both as a surveillance and as a diagnostic tool when invasive aspergillosis was suspected. It should be noted that the galactomannan assay had sensitivity-related limitations; lowering the cut-off value is expected to increase the diagnostic value for use in cases of invasive aspergillosis.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; beta-Glucans; Female; Galactose; Glucans; Hematologic Neoplasms; Humans; Limulus Test; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Sensitivity and Specificity

Recent case reports describe patients receiving piperacillin-tazobactam who were found to have circulating galactomannan detected by the double sandwich enzyme-linked immunosorbent assay (ELISA) system, leading to the false presumption of invasive aspergillosis. Since this property of piperacillin-tazobactam and galactomannan ELISA is not well understood, we investigated the in vitro, in vivo, and clinical properties of this interaction. Among the 12 reconstituted antibiotics representing four classes of antibacterial compounds that are commonly used in immunocompromised patients, piperacillin-tazobactam expressed a distinctively high level of galactomannan antigen in vitro (P = 0.001). After intravenous infusion of piperacillin-tazobactam into rabbits, the serum galactomannan index (GMI) in vivo changed significantly (P = 0.0007) from a preinfusion mean baseline value of 0.27 to a mean GMI of 0.83 by 30 min to slowly decline to a mean GMI of 0.44 24 h later. Repeated administration of piperacillin-tazobactam over 7 days resulted in accumulation of circulating galactomannan to a mean peak GMI of 1.31 and a nadir of 0.53. Further studies revealed that the antigen reached a steady state by the third day of administration of piperacillin-tazobactam. Twenty-six hospitalized patients with no evidence of invasive aspergillosis who were receiving antibiotics and ten healthy blood bank donors were studied for expression of circulating galactomannan. Patients (n = 13) receiving piperacillin-tazobactam had significantly greater mean serum GMI values (0.74 +/- 0.14) compared to patients (n = 13) receiving other antibiotics (0.14 +/- 0.08) and compared to healthy blood bank donors (0.14 +/- 0.06) (P < 0.001). Five (38.5%) of thirteen patients receiving piperacillin-tazobactam had serum GMI values > 0.5 compared to none of thirteen subjects receiving other antibiotics (P = 0.039) and to none of ten healthy blood bank donors (P = 0.046). These data demonstrate that among antibiotics that are commonly used in immunocompromised patients, only piperacillin-tazobactam contains significant amounts of galactomannan antigen in vitro, that in animals receiving piperacillin-tazobactam circulating galactomannan antigen accumulates in vivo to significantly increased and sustained levels, and that some but not all patients receiving this antibiotic will demonstrate circulating galactomannan above the threshold considered positive for invasive aspergillosis by the recently licensed

Topics: Animals; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Mannans; Penicillanic Acid; Piperacillin; Piperacillin, Tazobactam Drug Combination; Rabbits

Positive Platelia Aspergillus test results were observed in consecutive serum samples from an immunocompromised host during amoxicillin-clavulanic acid treatment, and a correlation between plasmatic amoxicillin concentration and galactomannan optical density index was observed. Amoxicillin-clavulanic acid vials tested positive for galactomannan but were negative for Aspergillus DNA.

Topics: Amoxicillin-Potassium Clavulanate Combination; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Immunocompromised Host; Mannans; Middle Aged; Reagent Kits, Diagnostic

Invasive pulmonary aspergillosis (IPA) is frequent and often fatal in hematopoietic stem cell transplant patients. Diagnosis requires microbiological or histopathologic demonstration of the organism in tissues; however, cultivation of Aspergillus species from respiratory secretions has low diagnostic sensitivity. Assays to detect Aspergillus antigen or DNA in bronchoalveolar lavage (BAL) fluid could facilitate earlier diagnosis, thereby guiding optimal therapy and obviating the need for additional costly and potentially morbid diagnostic evaluation. We evaluated the performance of a galactomannan enzyme immunoassay (GM EIA; Bio-Rad) by using a range of index cutoffs to define positivity and a quantitative PCR (qPCR) assay for the detection of Aspergillus species from BAL samples of patients with proven and probable IPA (case patients; n = 49) and without IPA (control patients; n = 50). The sensitivity of the GM EIA was 61% with an index cutoff of 1.0 and 76% with an index cutoff of 0.5; the corresponding specificities were 98 and 94%, respectively. The sensitivity and specificity of qPCR assay were 67 and 100%, respectively. The sensitivity with 22 culture-negative BAL specimens from patients with IPA was 41% for GM EIA with an index cutoff of 1.0, 59% for GM EIA with an index cutoff of 0.5, and 36% for qPCR assay. GM EIA indices and DNA quantities corresponded to BAL fungal burdens, with culture-positive samples having larger amounts of antigen and DNA compared to culture-negative samples. GM EIA and qPCR assay add to the sensitivity of BAL for diagnosing IPA in high-risk patients, with excellent specificity. Adjunctive use of these tests may reduce dependence on invasive diagnostic procedures.

Topics: Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Immunoenzyme Techniques; Lung Diseases, Fungal; Mannans; Polymerase Chain Reaction; Sensitivity and Specificity

Topics: Acute Disease; Amphotericin B; Antifungal Agents; Aspergillosis; Bone Marrow Transplantation; Bronchoalveolar Lavage Fluid; Child; Enzyme-Linked Immunosorbent Assay; Fatal Outcome; Galactose; Humans; Leukemia, Myeloid; Lung Diseases, Fungal; Male; Mannans; Postoperative Complications

The effectiveness of galactomannan detection with the Platelia test was evaluated in a prospective study of 3,327 sera from 807 patients. The specificity was 99.6% (748 of 751 cases). For the groups of patients with proven and probable invasive aspergillosis, the sensitivity was 50.0% (17 of 34 cases). The disappointing sensitivity associated with the presence of rare false-positive cases underlines the limits of this test.

Topics: Aged; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Child; False Positive Reactions; Female; Galactose; Humans; Male; Mannans; Mycology; Prospective Studies; Sensitivity and Specificity

Aspergillus fumigatus is the most common aetiological fungus responsible for human pulmonary aspergilloses. This study investigated the primary contact between Langerhans cells (LC), corresponding to dendritic cells present in pulmonary mucosa and live conidia of A. fumigatus. LC play a key role in antigen presentation for initiation of the primary T cell response. In vitro-generated LC (iLC) were differentiated from cultured human cord blood CD34+ cells and incubated at 4 degrees C or 37 degrees C with fluorescein-isothiocyanate (FITC)-stained conidia or control latex beads. In vitro, conidia were shown by microscopy and cytometry to adhere to iLC in a dose- and time-dependent manner. This adhesion was not limited to iLC because interstitial dendritic and other cells also fluoresced in the presence of conidia-FITC. A lectin other than mannose receptor-type lectin was demonstrated to be responsible of conidial binding. Inhibition of binding was observed with heterologous galactomannan and EDTA, indicating a C-lectin-like receptor with galactomannan structure specificity. After binding only a few conidia were internalized in acidic vesicles, as indicated by the cessation of conidial fluorescence. Conidial binding was followed by activation and maturation of iLC, suggesting that LC present in the lung may play a role in cellular host defence against aspergilloses.

Topics: Aspergillosis; Aspergillus fumigatus; Cell Adhesion; Edetic Acid; Flow Cytometry; Galactose; Humans; Langerhans Cells; Lectins; Lung; Lung Diseases, Fungal; Mannans; Microscopy, Fluorescence; Monensin; Mucous Membrane; Phagocytosis; Protein Binding

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; beta-Lactamase Inhibitors; Cross Reactions; Drug Therapy, Combination; False Positive Reactions; Galactose; Humans; Mannans; Penicillanic Acid; Piperacillin; Tazobactam

The diagnosis of invasive aspergillosis in neutropenic individuals is difficult and lengthy since non-invasive diagnostic tests lack sensitivity and specificity. The diagnosis of invasive aspergillosis in 154 prolonged neutropenic patients was prospectively bi-weekly validated by screening circulating galactomannan. The global sensitivity was 73% and specificity was 96%. The positive and negative predictive values were 73% and 98% respectively. False positive reactions occurred at a rate of 2%. Antigenemia was detected before clinical suspicion of invasive aspergillosis (median, 6 days before) in 30% of patients and anticipated the onset of radiologic signs 9 days in 60% of patients.. the prospective screening of galactomannan is a sensitive and non-invasive tool for early diagnosis of invasive aspergillosis in high-risk adult hematology patients.

Topics: Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Biomarkers; Bone Marrow Transplantation; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Fungemia; Galactose; Hematologic Neoplasms; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Neutropenia; Predictive Value of Tests; Prospective Studies; Risk; Sensitivity and Specificity; Time Factors

False-positive tests for Aspergillus galactomannan have been reported in neutropenic patients. We failed to detect any circulating antigen during the 2 weeks following allogeneic haematopoietic stem cell transplantation of 12 patients who had severe mucositis but were unable to eat.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; False Positive Reactions; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Parenteral Nutrition, Total

Screening for galactomannan (GM) has been adopted by many European centers as part of the management plan for allogeneic stem cell transplant recipients. However, the temporal onset of GM antigenemia remains unknown. A series of allogeneic stem cell transplant recipients were monitored prospectively, and the relationship between antigenemia and other diagnostic triggers for initiation of antifungal therapy was analyzed. GM detection had a sensitivity of 94.4% and a specificity of 98.8%. Positive and negative predictive values were 94.4% and 98.8%, respectively. This statistical profile was better than that of other triggers, including unexplained fever, new pulmonary infiltrates, isolation of Aspergillus species, and abnormalities seen on computed tomography. Antigenemia preceded diagnosis on the basis of radiologic examination or Aspergillus isolation by 8 and 9 days in 80% and 88.8% of patients, respectively. Antigenemia preceded therapy in 83.3% of patients. Detection of GM was especially useful when patients were receiving steroid treatment or when coexisting conditions masked the diagnosis of invasive aspergillosis. Prospective screening for GM allows earlier diagnosis of aspergillosis than do conventional diagnostic criteria.

Topics: Adolescent; Adult; Age Distribution; Anemia, Aplastic; Aspergillosis; Biomarkers; Female; Galactose; Hematologic Neoplasms; Humans; Male; Mannans; Middle Aged; Neoplasms; Stem Cell Transplantation; Survival Analysis; Time Factors; Transplantation, Homologous

Invasive aspergillosis is a serious problem for immunocompromised patients, especially if neutropenic. The diagnosis of this infection is complicated, since clinical symptoms are often similar to those of other fungal diseases. The chance of detecting the presence of a specific antigen in the serum could confirm the suspected clinical diagnosis and. perhaps, be useful for the follow-up of the patient. The Medical Mycology Committee of the Associazione Microbiologi Clinici Italiani (AMCLI) decided to evaluate in a multicenter prospective study (from I November 1998 to 28 February 1999) the performance of the Platelia Aspergillus Kit (Bio-Rad) for the detection of Aspergillus galactomannan in human serum. The enrolled patients included various groups of immunosuppressed patients (mostly neutropenic). Blood samples were drawn at the time of enrollment. This decision was based upon a clinical diagnosis of probable aspergillosis (antibiotic non-responsive fever for at least 96 hours, cough, hemophthosis and positive chest X-ray). Additional blood samples were drawn on days 3, 6, 9, 12, 15 and 21. Culture and histopathologic examinations were performed according to the individual laboratory workflow. For each patient the laboratory filled a form with all the available clinical information, to create a database on which to evaluate the results of the test. During the study, 187 patients with various kinds of immunosuppression were enrolled. A total of 256 sera were tested: for 117 patients (62.6%) only the basal sample was tested, whereas for the 70 symptomatic patients (37.4%) multiple specimens (range: 1-6) were tested. The results allowed the laboratories to exclude (68.6%) or confirm (31.5%: confirmed and/or probable) the clinical diagnosis of invasive aspergillosis; 4 cases remained undetermined. Based on the results of this study, it seems that the use of this test should be limited to those patients with clinical symptoms of aspergillosis.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Female; Galactose; Humans; Immunocompromised Host; Male; Mannans; Multicenter Studies as Topic; Neutropenia

The antifungal efficacy, pharmacokinetics, and safety of caspofungin (CAS) were investigated in the treatment and prophylaxis of invasive pulmonary aspergillosis due to Aspergillus fumigatus in persistently neutropenic rabbits. Antifungal therapy consisted of 1, 3, or 6 mg of CAS/kg of body weight/day (CAS1, CAS3, and CAS6, respectively) or 1 mg of deoxycholate amphotericin B (AMB)/kg/day intravenously for 12 days starting 24 h after endotracheal inoculation. Prophylaxis (CAS1) was initiated 4 days before endotracheal inoculation. Rabbits treated with CAS had significant improvement in survival and reduction in organism-mediated pulmonary injury (OMPI) measured by pulmonary infarct score and total lung weight (P < 0.01). However, animals treated with CAS demonstrated a paradoxical trend toward increased residual fungal burden (log CFU per gram) and increased serum galactomannan antigen index (GMI) despite improved survival. Rabbits receiving prophylactic CAS1 also showed significant improvement in survival and reduction in OMPI (P < 0.01), but there was no effect on residual fungal burden. In vitro tetrazolium salt hyphal damage assays and histologic studies demonstrated that CAS had concentration- and dose-dependent effects on hyphal structural integrity. In parallel with a decline in GMI, AMB significantly reduced the pulmonary tissue burden of A. fumigatus (P < or = 0.01). The CAS1, CAS3, and CAS6 dose regimens demonstrated dose-proportional exposure and maintained drug levels in plasma above the MIC for the entire 24-h dosing interval at doses that were > or =3 mg/kg/day. As serial galactomannan antigen levels may be used for therapeutic monitoring, one should be aware that profoundly neutropenic patients receiving echinocandins for aspergillosis might have persistent galactomannan antigenemia despite clinical improvement. CAS improved survival, reduced pulmonary injury, and caused dose-dependent hyphal damage but with no reduction in residual fungal burden or galactomannan antigenemia in persistently neutropenic rabbits with invasive pulmonary aspergillosis.

Topics: Animals; Anti-Bacterial Agents; Antibiotic Prophylaxis; Antifungal Agents; Aspergillosis; Aspergillus; Caspofungin; Disease Models, Animal; Echinocandins; Female; Galactose; Lipopeptides; Lung Diseases, Fungal; Mannans; Microbial Sensitivity Tests; Neutropenia; Peptides; Peptides, Cyclic; Rabbits; Treatment Outcome

The value of scintigraphic imaging using 99mTc labeled poly(ethyleneglycol) (PEG) -liposomes for detecting invasive pulmonary aspergillosis at different stages of the disease was investigated in a rat model. At 24, 48, 72, 120 and 168 h after fungal inoculation scintigraphic images were obtained and biodistribution of the radiolabel was determined. Findings were compared with serum galactomannan detection and other parameters of progression of fungal infection. At 48 h liposomal uptake in the infected left lung was increased significantly and 82% of the scintigraphic images was assessed positive. Serum galactomannan was only detected at 72 h and later. Liposomal uptake in the infected left lung increased over time and was significantly correlated with both the size of the pulmonary hemorrhagic lesion and the levels of circulating galactomannan. It was concluded that scintigraphic imaging using 99mTc-PEG-liposomes allows early detection of invasive pulmonary aspergillosis in this model, and that liposomal uptake in the infected lung was strongly associated with the severity of the disease.

Topics: Animals; Aspergillosis; Disease Progression; Female; Galactose; Kidney; Liposomes; Liver; Lung; Lung Diseases, Fungal; Mannans; Neutropenia; Polyethylene Glycols; Radionuclide Imaging; Radiopharmaceuticals; Rats; Spleen; Technetium; Tissue Distribution

The kinetics of serum Aspergillus galactomannan, as determined by enzyme-linked immunosorbent assay, was examined in 37 allogeneic stem cell transplant (SCT) recipients treated for invasive aspergillosis (IA). Fifty-eight periods of response ("response episodes") were evaluated. There were 42 response episodes that were considered "treatment failures" and 16 that were considered "good" (that is, complete or partial) responses. At baseline (the first day of each new response episode), the patients who experienced treatment failure and those who had good responses did not differ significantly with regard to median galactomannan index (GMI) value. Thereafter, GMI values significantly increased in the treatment failure group, whereas no significant changes were observed in the good response group (P=.002). An increase in the GMI value of 1.0 over the baseline value during the first week of observation was predictive of treatment failure with a sensitivity of 44%, a specificity of 87%, and a positive predictive value of 94%. We conclude that serial determination of serum GMI values is a useful tool for assessing prognosis of IA in allogeneic SCT recipients during treatment.

Topics: Adolescent; Adult; Antifungal Agents; Aspergillosis; Biomarkers; Child; Disease Progression; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Outcome Assessment, Health Care; Predictive Value of Tests; Prognosis; Sensitivity and Specificity; Transplantation, Homologous

Topics: Animals; Antifungal Agents; Aspergillosis; Food Contamination; Galactose; Humans; Infant; Male; Mannans; Milk

In common with a proportion of patients with invasive pulmonary aspergillosis (IPA), the efficacy of AmBisome treatment regimens in our rat model remains suboptimal. To investigate whether this might be the result of initially low antifungal activity of amphotericin B at the site of infection when administered in the liposomal form, Fungizone was added to AmBisome at the start of treatment. Groups of granulocytopenic rats with left-sided IPA received 10 day treatment regimens with either AmBisome 10 mg/kg/day (n = 25) or AmBisome 10 mg/kg/day combined with a single dose of Fungizone 1 mg/kg at day 1 (n = 27). Parameters of treatment response included survival, serum galactomannan (GM), size and quality of pulmonary macroscopic lesions, lung weight, viable fungal counts (cfu) and chitin content of the infected lung, and extra-pulmonary disseminated fungal infection. In a separate experiment the significance of early start of treatment to obtain therapeutic efficacy was investigated. Compared with untreated controls, both treatment regimens showed a significant increase in survival and change in parameters of fungal infection except left lung cfu. The combination treatment showed a significant increase in survival compared with AmBisome monotherapy (P = 0.02) and a significant decrease in left lung chitin content (P = 0.03). Differences in circulating GM concentrations between the two treatment regimes approached significance (P = 0.06). Delay in the start of treatment from 16 to 24 h after fungal inoculation resulted in a significant decrease in therapeutic efficacy (P = 0.02). It is concluded that the efficacy of AmBisome therapy can be enhanced by the addition of Fungizone at the start of treatment. This is probably a result of active amphotericin B being immediately available in the lung at the start of treatment.

Topics: Agranulocytosis; Amphotericin B; Animals; Antifungal Agents; Aspergillosis; Aspergillosis, Allergic Bronchopulmonary; Biomarkers; Chitin; Drug Therapy, Combination; Female; Galactose; Lung; Mannans; Rats; Survival Analysis; Time Factors

To improve the diagnosis of invasive aspergillosis (IA), we developed a LightCycler PCR assay targeted to Aspergillus fumigatus and A. flavus mitochondrial DNA. To avoid contamination, fully automated nucleic acid extraction with the MagNA Pure LC apparatus was used. The linearity of the results was achieved over a 6-log range of input A. fumigatus DNA, from 0.3 ng to 3 fg/10 microl of water. We retrospectively compared the LightCycler PCR and an enzyme-linked immunosorbent assay for the detection of galactomannan (GM) in serum from 14 patients with IA. The GM assay was more frequently positive (57 of 109; 52%) than the PCR assay (49 of 109; 45%). The LightCycler PCR assay, combined with automated DNA extraction, could be used in association with the GM assay to improve the reliability of IA diagnosis.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus flavus; Aspergillus fumigatus; Automation; DNA, Fungal; DNA, Mitochondrial; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction

Invasive aspergillosis (IA) is a well recognized, life-threatening infection in neutropenic patients and stem cell transplantation recipients. Early diagnosis is important to achieve the best outcome for these patients; however, definite proof often is difficult to obtain due to counterindicated invasive procedures.. This study evaluated the specificity and sensitivity of the detection of galactomannan (GM) for the diagnostic and prediction of IA in 347 children from the Pediatric Hematology Service and 450 patients from the Bone Marrow Transplantation Unit at the Hôpital Saint-Louis in Paris. Serial screening of Aspergillus GM circulating antigen was evaluated using a double sandwich ELISA assay (Platelia Aspergillus) on 6209 sera. Among the patients studied, 53 presented with confirmed IA (n = 27 patients) or probable IA (n = 26 patients).. Antigen was detected on at least two sequential sera in 48 of 53 patients, with a sensitivity of 90.6%. GM antigenemia was detected before the onset of radiologic signs in 31 of 48 patients (64.6%), with a mean of -8.4 days, and before clinical symptoms in 18 of 48 patients (39.6%), with a mean of -6.9 days. In patients without IA, 44 of 744 had positive antigenemia, resulting in a specificity of 94%. False positive results could not be related to the presence of a concurrent mucositis.. This large, prospective study allowed the authors to define better the conditions for the use of GM immunocapture ELISA in surveying patients who are at high risk for IA. The presence of antigen has a good diagnostic value mainly when there is an increase in the titer on two consecutive sera samples. A repeated negative result is a strong argument against the diagnosis of IA; however, an awareness of the possibility of unexplained false negative results is important.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Child; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Immunocompromised Host; Male; Mannans; Middle Aged; Prospective Studies; Sensitivity and Specificity

The antifungal efficacy, safety, and pharmacokinetics of posaconazole (SCH 56592) (POC) were investigated in treatment and prophylaxis of primary pulmonary aspergillosis due to Aspergillus fumigatus in persistently neutropenic rabbits. Antifungal therapy consisted of POC at 2, 6, and 20 mg/kg of body weight per os; itraconazole (ITC) at 2, 6, and 20 mg/kg per os; or amphotericin B (AMB) at 1 mg/kg intravenously. Rabbits treated with POC showed a significant improvement in survival and significant reductions in pulmonary infarct scores, total lung weights, numbers of pulmonary CFU per gram, numbers of computerized-tomography-monitored pulmonary lesions, and levels of galactomannan antigenemia. AMB and POC had comparable therapeutic efficacies by all parameters. By comparison, animals treated with ITC had no significant changes in outcome variables in comparison to those of untreated controls (UC). Rabbits receiving prophylactic POC at all dosages showed a significant reduction in infarct scores, total lung weights, and organism clearance from lung tissue in comparison to results for UC (P < 0.01). There was dosage-dependent microbiological clearance of A. fumigatus from lung tissue in response to POC. Serum creatinine levels were greater (P < 0.01) in AMB-treated animals than in UC and POC- or ITC-treated rabbits. There was no elevation of serum hepatic transaminase levels in POC- or ITC-treated rabbits. The pharmacokinetics of POC and ITC in plasma demonstrated dose dependency after multiple dosing. The 2-, 6-, and 20-mg/kg dosages of POC maintained plasma drug levels above the MICs for the entire 24-h dosing interval. In summary, POC at > or =6 mg/kg/day per os generated sustained concentrations in plasma of > or =1 microg/ml that were as effective in the treatment and prevention of invasive pulmonary aspergillosis as AMB at 1 mg/kg/day and more effective than cyclodextrin ITC at > or =6 mg/kg/day per os in persistently neutropenic rabbits.

Topics: Animals; Antibiotic Prophylaxis; Antifungal Agents; Aspergillosis; Aspergillus fumigatus; Disease Models, Animal; Female; Galactose; Itraconazole; Lung Diseases, Fungal; Mannans; Neutropenia; Rabbits; Treatment Outcome; Triazoles

The diagnosis of invasive aspergillosis (IA) in patients with hematologic disorders is not straightforward; lack of sensitive and specific noninvasive diagnostic tests remains a major obstacle for establishing a precise diagnosis. In a series of 362 consecutive high-risk treatment episodes that were stratified according to the probability of IA based on recently accepted case definition sets, the potential for diagnosis of serial screening for circulating galactomannan (GM), a major aspergillar cell wall constituent was validated. After incorporating postmortem findings to allow a more accurate final analysis, this approach proved to have a sensitivity of 89.7% and a specificity of 98.1%. The positive and negative predictive values equaled 87.5% and 98.4%, respectively. False-positive reactions occurred at a rate of 14%, although this figure might be overestimated due to diagnostic uncertainty. More or less stringent criteria of estimation could highly influence sensitivity, which ranged from 100% to 42%; the impact on other test statistics was far less dramatic. All proven cases of IA, including 23 cases confirmed after autopsy only, had been detected before death, although serial sampling appeared to be necessary to maximize detection. The excellent sensitivity and negative predictive value makes this approach suitable for clinical decision making. Unfortunately, given the species-specificity of the assay, some emerging non-Aspergillus mycoses were not detected. In conclusion, serial screening for GM, complemented by appropriate imaging techniques, is a sensitive and noninvasive tool for the early diagnosis of IA in high-risk adult hematology patients.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aspergillosis; Autopsy; Biomarkers; Cohort Studies; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Neoplasms; Hematopoietic Stem Cell Transplantation; Humans; Male; Mannans; Middle Aged; Neutropenia; Predictive Value of Tests; Prospective Studies; Sensitivity and Specificity

Topics: Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Chronic Disease; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Graft vs Host Disease; Humans; Mannans; Transplantation, Homologous

The sensitivity of a sandwich enzyme-linked immunosorbent assay (ELISA) for detecting Aspergillus galactomannan was evaluated with 66 serum samples and 113 specimens of the respiratory tract obtained from 52 patients with pulmonary diseases. The patients were divided into five groups: proven invasive pulmonary aspergillosis (IPA) (five patients), probable IPA (seven patients), Aspergillus colonization (eight patients) or unlikely Aspergillus infection (27 patients). Another five patients with doubtful diagnostic test results are discussed in detail. The results of the Platelia Aspergillus ELISA (Sanofi Pasteur, Freiburg, Germany) in testing specimens of the respiratory tract were 90% sensitivity in proven (serum 38%), 60% in probable (serum 37%) and 71% in Aspergillus colonization (serum 0%). Furthermore, 85% of the Aspergillus spp. from positive cultures of specimens of the respiratory tract were also detected in the ELISA. A total of 57% of the culture negative specimens of patients with a least one positive culture or proven aspergillosis in a series of specimens were positive in the ELISA.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Sensitivity and Specificity

Intravenous (i.v.) infection of immunocompetent mice with Aspergillus fumigatus was used to investigate the ability of a commercial galactomannan enzyme-linked immunosorbent assay (ELISA) to monitor the course of organ infection after dissemination. The test detected 100% of the fungemias which occurred for up to 5 days after infection. When blood-cultures became negative but there was a high load of fungi in the parenchymal organs and a positive culture from the brain, the ELISA was again positive in all animals. However, when blood cultures as well as brain cultures were negative and lower amounts of fungi demarcated by immune cells were present in the liver and kidneys which was the case between day 5 and 30 of infection, the test was negative in most of the animals. Therefore, the test was excellent for detection of early i.v. infection with Aspergillus fumigatus but not suited for detection of limited organ infection in immunocompetent mice.

Topics: Animals; Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Fungemia; Galactose; Injections, Intravenous; Mannans; Mice; Mice, Inbred BALB C

Pulmonary invasive aspergillosis is frequently difficult to diagnose. In particular, the value of the cultivation of Aspergillus and the Aspergillus galactomannan antigen detection (Pastorex) in bronchoscopically acquired material (bronchoalveolar lavage = BAL, bronchial lavage = BL) in the course of diagnosing this mycosis is viewed controversially. Between January 1996 and September 1999, we obtained 114 positive results in 100 bronchoscopically aquired specimens from a total of 69 patients. 59 of the 69 patients were immunosuppressed, 42 suffered from pulmonary aspergillosis and 38 suffered from invasive pulmonary aspergillosis. The positive prediction rate for a positive result with regard to pulmonary aspergillosis in bronchoscopically acquired material was approximately 61%. Cultivation of Aspergillus was more successful in BAL, and the Aspergillus antigen detection was more successful in BL.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage; Galactose; Humans; Lung Diseases, Fungal; Mannans; Retrospective Studies

Topics: Adult; Aged; Aspergillosis; Aspergillus; Culture Media; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Latex Fixation Tests; Male; Mannans; Middle Aged; Rhinitis; Sinusitis

Primary intestinal invasive aspergillosis is rarely reported in leukaemic patients. We describe a case of jejunal invasive aspergillosis in the setting of aplasia following chemotherapy for acute myeloid leukaemia. The diagnosis was confirmed by biopsy obtained during surgery and our polymerase chain reaction (PCR) test confirmed Aspergillus flavus as the fungus responsible. This patient had high levels of circulating galactomannan, an antigen secreted by Aspergillus sp., in serum. The ELISA test for galactomannan has been developed to improve the diagnosis of invasive aspergillosis but presents a 5-15% false positive rate. We suggest that some false positive results might be due to non-respiratory invasive aspergillosis, the usual localization of invasive aspergillosis. Our PCR test was also positive in serum. In case of positive results in serum with antigen and/or PCR tests without respiratory symptoms, the intestinal localizations should be investigated.

Topics: Aspergillosis; Aspergillus flavus; Biopsy; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Fatal Outcome; Female; Galactose; Humans; Intestinal Diseases; Jejunum; Leukemia, Myeloid, Acute; Mannans; Middle Aged; Polymerase Chain Reaction

Diagnosis of canine aspergillosis is difficult using currently available methods. It often passes unnoticed or is diagnosed in the later phases of the disease. We developed an ELISA technique to detect anti-Aspergillus antibodies in canine serum using an Aspergillus antigenic mycelial extract, which could then be used for the diagnosis of canine aspergillosis. We used a cut-off of X + 3SD obtained from 20 control sera. The test was performed on 46 dogs with lesions indicating possible aspergillosis and gave nine positive results: one systemic mycosis, two discospondylitis, one uveitis, two bronchopulmonary processes and three rhinitis. We compared this methodology with the PLATELIA technique in the follow-up of the affected dogs, obtaining the same limitations as in the diagnosis of human aspergillosis. We consider our ELISA technique using sera samples a speedy, safe and reliable method which enables us to follow up the evolution of the disease and the efficacy of the therapy chosen. A definitive diagnosis must still take into account the results of other tests such as clinical examination, radiographic studies, endoscopy and biopsy.

Topics: Animals; Antibodies, Fungal; Aspergillosis; Aspergillus; Dog Diseases; Dogs; Enzyme-Linked Immunosorbent Assay; Galactose; Immunoglobulin G; Mannans; Reproducibility of Results; Sensitivity and Specificity

We have developed an inhibition enzyme immunoassay (inhibition-EIA) to monitor for the occurrence of invasive aspergillosis (IA) in sera from 45 immunocompromised (IC) patients. The test uses rabbit polyclonal antibodies and a mixture of components from Aspergillus fumigatus, containing three predominant antigens with molecular weights of 18,000, 33,000, and 56,000. Circulating antigens were found in five of seven proven cases of IA due to A. fumigatus. In two of the five positive cases, antigenemia was detected with inhibition-EIA earlier than with X ray or other biological methods. No antigens were detected in the sera from two patients with proven IA due to Aspergillus flavus and Aspergillus terreus nor in the sera from four patients with probable IA. Circulating antigens were not detected in the control group, composed of 30 healthy adult blood donors. Four of the 32 at-risk patients examined, though they displayed no definite evidence of IA, gave a positive result in this test. The sensitivity, specificity, and positive predictive value of inhibition-EIA were 71.4, 94.4, and 71.2%, respectively. The data were compared with those obtained by a latex agglutination assay of galactomannan (GM) that was positive in only one patient with probable IA. The higher sensitivity obtained by inhibition-EIA may well be due to its ability to detect circulating antigens other than GM in the sera of IC patients with IA. Detecting these antigens may improve the diagnosis of IA, as they may serve as markers of this infection.

Topics: Antigens, Fungal; Aspergillosis; Galactose; Hot Temperature; Humans; Immunocompromised Host; Immunoenzyme Techniques; Latex Fixation Tests; Mannans; Risk Factors; Time Factors

Two diagnostic tests, an Aspergillus-specific PCR and an enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of galactomannan, were compared for diagnosing and monitoring invasive pulmonary aspergillosis. Persistently neutropenic rats with left-sided invasive pulmonary aspergillosis were sacrificed at regular intervals after inoculation. Blood samples and bronchoalveolar lavage (BAL) fluid were cultured and tested by PCR as well as by ELISA. Disseminated fungal infection in extrapulmonary organs was determined. The sensitivity of the ELISA was higher than that of the PCR on all days of measurements, in both blood and BAL fluid. Positive PCR or ELISA results in blood were not significantly associated with disseminated fungal infection. Serial testing in a separate group of rats showed consistently increasing concentrations of circulating galactomannan during the course of disease, while a positive PCR could be followed by negative results. The concentration of galactomannan was highly predictive for the time of survival (P < 0.0001). It was concluded that, in this model, quantitative galactomannan detection is superior to PCR in diagnosing and monitoring invasive pulmonary aspergillosis.

Topics: Animals; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Disease Models, Animal; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Lung Diseases, Fungal; Mannans; Polymerase Chain Reaction; Rats; Reproducibility of Results

Topics: Antigens, Fungal; Aspergillosis; Galactose; Hematologic Diseases; Humans; Mannans; Risk Factors

We report a patient with chronic granulomatous disease who developed invasive pulmonary aspergillosis and a subphrenic abscess. During treatment, high levels of Aspergillus antigen were detected in the abscess, but circulating antigen and Aspergillus DNA were undetectable in the serum.

Topics: Antifungal Agents; Aspergillosis; Aspergillus; Biopsy, Needle; Child, Preschool; Diagnostic Errors; DNA, Fungal; Galactose; Granulomatous Disease, Chronic; Humans; Itraconazole; Lung Abscess; Lung Diseases, Fungal; Male; Mannans; Pyrimidines; Triazoles; Voriconazole

The incidence of invasive fungal infections is increasing in patients with hematological malignancies. Invasive aspergillosis is one of the most frequently encountered infections with a high mortality rate. New diagnostic tests for invasive aspergillosis such as the detection of Aspergillus galactomannan antigen by a sandwich enzyme-linked immunosorbent assay (ELISA) have recently been described. The objective of this study was to evaluate this assay as a potential surrogate for invasive procedures used to diagnose IA.. We analyzed the performance of a commercially available ELISA test which we routinely use for the surveillance of galactomannan antigenemia in patients with hematological malignancies experiencing chemotherapy-induced prolonged neutropenia (ANC < 500/mm(3) for more than 7 days). Serum samples were collected on a weekly basis. Test positivity was defined in accordance with the manufacturer's recommendations.. Over the 2 year study period, we analyzed 507 samples obtained during 193 neutropenic episodes from 135 patients. Ten, six and two patients were considered to have proven, probable or possible invasive aspergillosis, respectively, based on clinical, radiological or microbiological data. Forty-four positive (Index>1.5) and 26 'undetermined' (1.5 > Index > 1.0) test results were observed in 17 and ten patients respectively. All invasive aspergillosis cases had at least a positive or an undetermined test result. Only one positive and one undetermined result were found in two patients before the onset of clinical or radiological signs suggesting invasive aspergillosis. Sensitivity was 69% and specificity 96% if only positive results are considered; when 'undetermined' test results were combined with positive results, sensitivity attained 100% and specificity 92% suggesting that the cutoff value for positivity can be lowered from 1.5 to 1.0.. Although the ELISA test did not appear to play a role in the early diagnosis of invasive aspergillosis and in the anticipation of antifungal therapy in our experience, it clarifies the diagnosis of infection in probable or possible invasive aspergillosis especially when the cutoff value is lowered and is useful for monitoring patients receiving specific therapy.

Topics: Adolescent; Adult; Aged; Antigens, Fungal; Antineoplastic Agents; Aspergillosis; Aspergillosis, Allergic Bronchopulmonary; Aspergillus; Child; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Neoplasms; Humans; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Myeloid, Acute; Lymphoma, Non-Hodgkin; Male; Mannans; Middle Aged; Neutropenia

We evaluated the usefulness of PCR and antigen detection for the diagnosis of pulmonary aspergillosis. Forty-four serum samples from patients with pulmonary aspergillosis (33 with pulmonary aspergilloma, 4 with allergic bronchopulmonary aspergillosis, 4 with invasive pulmonary aspergillosis, and 3 with aspergillus pyothorax) were used in this study. PCR detection of Aspergillus DNA in serum samples was successful in 39 patients. Galactomannan antigen was detected by sandwich enzyme-linked immunosorbent assay in 25 patients and by latex agglutination test in 13 patients. Detection of Aspergillus DNA in serum samples by nested PCR had the highest sensitivity of the three methods tested for the diagnosis of pulmonary aspergillosis.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Lung Diseases, Fungal; Mannans; Microbiological Techniques; Polymerase Chain Reaction; Sensitivity and Specificity; Serologic Tests

The performance of antibody detection, antigen detection, and Aspergillus genus-specific PCR for diagnosing Aspergillus meningitis was investigated with 26 cerebrospinal fluid (CSF) samples obtained from a single patient with proven infection caused by Aspergillus fumigatus. Immunoglobulin G antibodies directed against Aspergillus were not detected by enzyme-linked immunosorbent assay in CSF or serum. The antigen galactomannan was detected in the CSF 45 days before a culture became positive, and Aspergillus DNA was detected 4 days prior to culture. Decline of the galactomannan antigen titer in the CSF during treatment with intravenous and intraventricular amphotericin B and intravenous voriconazole corresponded with the clinical response to treatment.

Topics: Aged; Amphotericin B; Antibodies, Fungal; Antifungal Agents; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; DNA, Fungal; Drug Resistance, Microbial; Female; Galactose; Humans; Mannans; Meningitis, Fungal; Mycology; Polymerase Chain Reaction; Pyrimidines; Time Factors; Triazoles; Voriconazole

We compared the usefulness of a polymerase chain reaction (PCR) assay for the early diagnosis of invasive pulmonary aspergillosis with the serodiagnosis of sufficient concentrations of galactomannan using the same serum samples. A patient was treated with prednisolone for the management of hepatitis. Computed tomography (CT) scan of the chest showed the nodular shadow with a cavity containing a clear fungus ball. DNA of Aspergillus spp. from a serum sample was detected and using the same serum sample, both latex agglutination and sandwich enzyme-linked immunosorbent assay (ELISA) of galactomannan were negative. PCR assay provides an early diagnosis of invasive pulmonary aspergillosis compared with ELISA of galactomannan.

Topics: Antibodies, Fungal; Aspergillosis; Aspergillus fumigatus; Bronchoalveolar Lavage Fluid; Diagnosis, Differential; DNA, Fungal; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Latex Fixation Tests; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Radiography, Thoracic; Tomography, X-Ray Computed

Efforts to improve the diagnosis of invasive aspergillosis (IA) have been directed towards the detection of fungal antigens, including galactomannan (GM). However, previous evaluations of GM detection have been hampered by a lack of proven cases of IA and by a nonserial study design. This prospective study assessed the diagnostic value of serial screening for circulating GM by using a recently developed sandwich enzyme-linked immunosorbent assay (ELISA) for prolonged-neutropenic and/or steroid-treated patients with hematological disorders. Serum GM levels were monitored twice weekly for 186 consecutive patients at increased risk for IA. The patients were stratified according to the likelihood of IA (proven, probable, possible, and no evidence of IA) by using stringent criteria. Proven IA was defined by characteristic histopathological findings together with a positive culture for Aspergillus species. Autopsy and culture from autopsy specimens was used to verify both positive and negative test results. A total of 2,172 serum samples were tested from 243 episodes (mean, 9 samples/episode). Based on the analysis of 71 patients with confirmed disease status (culture and histology), the sensitivity and specificity of serial GM monitoring were 92.6 and 95.4%, respectively. The positive predictive value was almost 93%, the negative predictive value was 95%, and the efficacy was 94%. False-positive reactions occurred at a rate of nearly 8%, although this figure might have been overestimated. Less than 1% of all tested sera were considered inconclusive. In more than half of the cases, antigenemia was detected before clinical suspicion of IA (median, 6 days before). Serial determination of serum GM by the sandwich ELISA technique is a sensitive tool for the diagnosis of IA in hematological patients at risk. This approach may substantially influence clinical management with regard to preemptive and empirical antifungal therapy.

Topics: Adolescent; Adult; Aged; Aspergillosis; Autopsy; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Hematologic Diseases; Humans; Infant; Male; Mannans; Middle Aged; Prospective Studies; Risk

The performance of two Aspergillus antigenemia systems, the sandwich enzyme-linked immunosorbent assay (ELISA), Platelia Aspergillus test, and the latex agglutination (LA), Pastorex Aspergillus test, in the diagnosis of invasive aspergillosis were compared by testing 364 serum samples from 22 bone marrow transplant (BMT) recipients. Sensitivity and specificity for the ELISA test were 60% and 82% respectively, vs 40% and 94% for the LA test. In the two patients found positive with both methods, the ELISA test became positive earlier than the LA test or remained positive after the LA test had become negative. These results encourage further evaluation of the Platelia Aspergillus test, to assess its role in the management of invasive aspergillosis in BMT patients.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Latex Fixation Tests; Lung Diseases, Fungal; Male; Mannans; Prospective Studies; Sensitivity and Specificity

The intra- and interlaboratory reproducibilities of a commercial sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of Aspergillus galactomannan in serum (Platelia Aspergillus; Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) were evaluated in six laboratories of university hospitals. Twenty serum samples were obtained from 12 neutropenic patients including 6 with invasive aspergillosis. These samples were blinded and sent to each center together with eight blinded ELISA-negative serum samples spiked with known concentrations of galactomannan. The centers were provided with ELISA microtiter plates from a single batch and a detailed protocol. Ten clinical samples showed ELISA reactivity, while 10 samples were ELISA negative. The mean coefficient of variation (CV) of the optical density values was 4.24% within a single assay and 25.6% between runs. The interassay CV of the ratios for the serum samples tested was 18.6%. Analysis of ordinal interpretation of the ELISA result (i.e., negative, gray zone, or positive) showed excellent reproducibility. Recalculation of the cutoff values for positive and negative samples suggested that the cutoff level recommended by the manufacturer could be lowered from 1.0 to 0.8 for negative samples and from 1.5 to 1.0 for positive samples. The intra- and interlaboratory reproducibilities were excellent when the ELISA results were interpreted as ordinal data, but considerable variation in optical density values and, to a lesser extent, in the ratios for the serum samples tested, was observed between runs. High assay variability was also found for serum samples spiked with known concentrations of galactomannan. Therefore, antigen titers in serum samples from a single patient, measured in different runs, should be compared with caution.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Galactose; Hospitals, University; Humans; Laboratories, Hospital; Mannans; Netherlands; Neutropenia; Observer Variation; Quality Control; Reproducibility of Results

To improve the diagnosis of invasive aspergillosis (IA), we retrospectively compared competitive polymerase chain reaction (PCR) and sandwich ELISA for detection of serum galactomannan (GM) antigen. We studied 281 serum samples collected weekly during the period at risk for IA from 41 selected hematology patients. Twenty-two patients had confirmed, probable, or suspected IA, according to clinical and mycologic data. Fifteen of them had positive GM titers (87 samples) and 12 had positive PCRs (20 samples). Nineteen of the 20 PCR-positive samples were also GM-positive. Of the 19 patients without IA (83 samples), one had 3 GM-false-positive samples. Neither test anticipated the initiation of antifungal therapy on the basis of clinical suspicion. Both tests were more likely to be positive before death. This study suggests that PCR on serum samples is not more sensitive than GM detection. However, PCR can improve the specificity of the GM test. Together, these noninvasive tests should improve the diagnosis of IA.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Child; Female; Galactose; Humans; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Retrospective Studies

A new sandwich antigen enzyme-linked immunosorbent assay (ELISA) for the detection of Aspergillus galactomannan in blood plasma was compared with the latex-agglutination test by investigation of six patients with histologically proven invasive aspergillosis. The ELISA gave positive results in all six patients while the latex test recognized 5/6 as positive. On an average the antigen ELISA gave positive results 19 days earlier than the latex test.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Female; Galactose; Humans; Latex Fixation Tests; Male; Mannans; Middle Aged; Retrospective Studies

With a view to improving the sensitivity of serological detection of Aspergillus galactomannan (GM), a europium-linked time-resolved fluoroimmunoassay was developed. This method was compared to an enzyme-linked immunosorbent assay using a peroxidase-conjugated detector antibody. No increase in the sensitivity of the detection of GM standards was seen with the europium-based fluoroimmunoassay.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; Europium; Fluorescent Antibody Technique; Galactose; Humans; Mannans; Sensitivity and Specificity

We compared PCR, galactomannan detection assay using a latex agglutination test and (1-->3)-beta-D-glucan detection assay in detecting infection in rats experimentally infected with Aspergillus fumigatus. On day 2 after inoculation, (1-->3)-beta-D-glucan and nested PCR were positive for 80%, while galactomannan detection assay was positive for 60%. In addition, the positive result of nested PCR (87.5%) was higher than those of galactomannan detection assay (75%) and (1-->3)-beta-D-glucan (71.4%) on day 3 after inoculation. The sensitivity of nested PCR was superior to those of galactomannan detection assay and (1-->3)-beta-D-glucan detection assay. The three diagnostic tests were compared with histopathological findings, and the sensitivity of three diagnostic tests was correlated with histopathological changes. In addition, the elevated levels of (1-->3)-beta-D-glucan paralleled the development and progression of pulmonary aspergillosis. Our results indicate that a combination of two or three of these tests seems to provide a rapid diagnosis of invasive aspergillosis and assist in the evaluation of the development and severity of invasive aspergillosis.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Galactose; Glucans; Latex Fixation Tests; Lung Diseases, Fungal; Male; Mannans; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley

Pulmonary aspergillosis is classified into invasive, saprophytic, and allergic forms. In this study, we evaluated the usefulness of PCR for differentiating between different forms of aspergillosis or in monitoring disease activity during treatment by detecting DNA specific for Aspergillus species in the serum. Nested PCR was used to detect Aspergillus DNA in the sera of 30 patients with various forms of pulmonary aspergillosis. The results were compared with those of latex agglutination tests for detecting galactomannan antigen. We also examined the serial changes in the results of nested PCR during and after treatment of a subgroup of patients with invasive pulmonary aspergillosis with amphotericin B. The highest proportion of positive nested PCR results were in patients with invasive aspergillosis (10 of 12; 83%), while patients with pulmonary aspergilloma had the lowest frequency of positive tests (1 of 9; 11%). These results suggested that the sensitivity of the nested PCR depends on the extent of invasion by Aspergillus species. Serial assays showed that the results of nested PCR became negative shortly after commencement of antifungal treatment and that such changes did not correlate with clinical responsiveness to treatment. Our results indicate the potential usefulness of nested PCR with serum samples for the diagnosis of invasive aspergillosis and the detection of a shift in the status of infection from a noninvasive type to invasive aspergillosis. However, the results of the nested PCR did not correlate with the response to antifungal treatment.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Fungal; Aspergillosis; Aspergillus; DNA, Fungal; Female; Galactose; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Polymerase Chain Reaction

The sensitivity of a sandwich enzyme-linked immunosorbent assay (ELISA) for detecting Aspergillus galactomannan was tested using 783 serum samples obtained from 247 patients (1-15 sera per patient) with severe underlying diseases (haematological malignancies or intensive care unit stay). We selected 146 serum samples from 50 patients for retesting. Serum samples were frozen after routine testing at -18 degrees C until retesting. All patients charts were checked for signs of Aspergillus infection, such as pneumonia or sinusitis. Adult patients were divided into four groups: proven (5), probable (6), suspected (8) or unlikely (25) Aspergillus infection. The results of Platelia ELISA were 100% in proven, 33% in probable and 50% in suspected Aspergillus infection. Patients with unlikely infection had no positive results with Platelia ELISA. Group 5 consists of six paediatric patients with prolonged ICU stay and a birth weight of 400-1320 g. In five out of six infants we found positive results with Platelia ELISA. All positive results in this group of patients are considered as false positive (83.3%).

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Enzyme-Linked Immunosorbent Assay; False Positive Reactions; Female; Galactose; Humans; Infant; Infant, Newborn; Infant, Premature; Infant, Premature, Diseases; Lung Diseases, Fungal; Male; Mannans; Middle Aged; Sensitivity and Specificity

The interaction between Aspergillus fumigatus conidia and murine macrophages of various origins was investigated. Cocultures were carried out between A. fumigatus strains and freshly isolated murine pulmonary alveolar macrophages or two murine macrophage cell-lines: murine alveolar cell-line MALU and murine astrocytoma cell-line J774. By measuring the variation of elastolytic activity in the coculture supernatants with two elastin substrates, we demonstrated that either viable or fixed A. fumigatus or C. albicans yeasts or nonspecific particles induced significant macrophage elastolytic activity. The effect of A. fumigatus supernatant or the purified A. fumigatus galactomannan suggested also the possible involvement of this polysaccharide in macrophage-protease gene expression, release, and activity in invasive aspergillosis. The effect of inhibitory compounds demonstrated the potential implication of a macrophagic metalloprotease and a macrophagic cysteine protease. RNA analysis allowed us to demonstrate the induction of expression of two macrophagic protease genes in stimulated macrophages. Two distinctive mechanisms appeared to be implicated in macrophage protease induction: nonspecific phagocytosis in the earliest times of the coculture and (or) specific galactomannan recognition after its gradual release by the mycelium.

Topics: Animals; Aspergillosis; Aspergillus fumigatus; Cell Adhesion; Cells, Cultured; Cysteine Endopeptidases; Elastin; Endopeptidases; Galactose; Gene Expression; Humans; Lung Diseases, Fungal; Macrophages, Alveolar; Mannans; Metalloendopeptidases; Mice; Mice, Inbred C57BL; Phagocytosis; Protease Inhibitors; RNA, Messenger; Spores, Fungal

The specificity of the Pastorex Aspergillus latex agglutination test for the diagnosis of manifest aspergillosis is hampered by the occurrence of false-positive results. In order to prove whether or not the false-positive reactions may be caused by the uptake of the soluble galactomannan antigen from the environment, the presence of the antigen was tested in foods, air samples, antibiotics for therapeutic use and faeces. Reactions of the Aspergillus latex agglutination test were found in 15 (79%) out of 19 samples of meals prepared in a hospital kitchen, in five out of six canned vegetables from a supermarket, in all of six samples of pasta and rice bought in health shops, in the faeces of four bone marrow transplant (BMT) recipients and of four healthy subjects and in one and two batches of the antibiotics co-amoxyclav and piperacillin respectively. The concentration of the antigen in faecal material was calculated to be in the range of 1.2-38.4 micrograms g-1. It is concluded that the faecal galactomannan antigen may reach the circulation in patients with dysfunction of the intestinal mucosal barrier, e.g. BMT recipients, thus leading to diagnostically false-positive antigenaemia.

Topics: Air Microbiology; Anti-Bacterial Agents; Antigens, Fungal; Aspergillosis; Aspergillus; Drug Contamination; False Positive Reactions; Feces; Food Microbiology; Galactose; Humans; Latex Fixation Tests; Mannans

Aspergillus antigenemia was followed up in 215 consecutively observed bone marrow transplant (BMT) patients over a period of two years, using both a latex agglutination test and a sandwich immunocapture enzyme immunoassay (EIA) with a rat antigalactomannan monoclonal antibody as capture and detector antibody. For each patient, sequential sera (3 to 20) were obtained before and after BMT. No positivity was observed before BMT. After BMT, the EIA and latex agglutination test were positive in 19 and 4 patients respectively of 25 patients with confirmed aspergillosis and 14 and 7 of 15 patients with probable aspergillosis. In 19 of 25 patients with confirmed aspergillosis and 9 of 15 patients with probable aspergillosis, the EIA was more sensitive and detected infection earlier than the latex test. In all positive cases, antigenemia rapidly increased in sequential samples and remained strongly positive. In 31 of 169 (19%) BMT patients without clinical signs of aspergillosis, the EIA was occasionally positive in samples taken within the first month after BMT, giving a specificity of 81% in these patients. In non-BMT patients suffering from other diseases (n = 77), the specificity was 98.7%. The overall positive and negative predictive values for the EIA were 54% and 95% respectively. These results favour the use of EIA for early diagnosis and monitoring of aspergillosis in BMT patients, although the predictive value of transient positivity remains to be ascertained.

Topics: Animals; Antibodies, Monoclonal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Female; Fungemia; Galactose; Humans; Immunocompromised Host; Immunoenzyme Techniques; Latex Fixation Tests; Male; Mannans; Predictive Value of Tests; Rats; Reproducibility of Results; Sensitivity and Specificity

During the course of extensive building activity in the vicinity of bone marrow transplantation wards, the patients were routinely screened for the occurrence of Aspergillus galactomannan antigen in serum. In 19 (6.7%) out of 285 patients, an antigenemia was detected. Eleven (58%) of the 19 antigenemic patients suffered from autopsy-proven or clinically suspected invasive aspergillosis. The yearly incidence of antigenemic patients differed significantly, ranging from 0% in the year without building activities to 20.9% in the year with major activities, particularly interior completion works and landscaping. It is concluded that Aspergillus antigen monitoring of bone marrow transplant recipients has a limited value for the diagnosis of manifest invasive aspergillosis. However, it it epidemiologically useful to assess the extent of intensive contact with aspergilli and to control the effectivity of preventive measures.

Topics: Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Galactose; Humans; Incidence; Mannans

The delay between the onset of invasive aspergillosis and the start of antifungal therapy is crucial for the patient's recovery. Early diagnosis is difficult in cancer patients through lack of precocious specific signs. We have investigated the clinical usefulness of circulating Aspergillus antigen monitoring in pediatric hematology patients with a new sensitive sandwich enzyme-linked immunosorbent assay.. A prospective study was conducted by assessing circulating galactomannan levels in high risk patients. Thirty-seven patients studied during an 18-month period were evaluated twice weekly during neutropenic phases with the sandwich enzyme-linked immunosorbent assay for serum Aspergillus galactomannan.. Twelve patients had one or more episodes of positive circulating galactomannan detection, 10 of whom developed presumptive invasive aspergillosis. The clinical and radiologic signs occurred at a mean of 13.4 days (range, 0 to 48) after circulating galactomannan detection and reversed in 6 patients treated with amphotericin B at the same time circulating galactomannan detection became negative. Reappearance of circulating galactomannan was observed during subsequent neutropenic periods in 3 patients.. The detection of galactomannan at concentrations as low as 1 ng/ml can be useful for the early initiation of antifungal therapy and monitoring treatment in clinically documented lung aspergillosis. This technique coupled with chest computed tomography could help to restrict the need of invasive diagnostic procedures in fragile patients.

Topics: Adolescent; Antigens, Fungal; Aspergillosis; Aspergillus; Child; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Follow-Up Studies; Galactose; Humans; Mannans; Opportunistic Infections; Predictive Value of Tests; Prospective Studies

Topics: Aspergillosis; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans

Foeto-placental infections are obtained when pregnant mice are challenged intravenously with conidial suspensions of Aspergillus fumigatus. This experimental model, which is used without any immuno-suppressive pretreatment, can be employed to screen for differences in foeto-placental infectivity of A. fumigatus strains when the number of colony-forming units (CFUs) in conidial suspensions used for infection is from 1 x 10(5) to 1 x 10(6). In the foeto-placental unit, hyphal growth was initiated at the periphery of the placental disc from which infection spread to the central parts of the placenta, the extrafoetal membranes and the foetus. Complement activation was noticed as a consequence of pregnancy and conidial inoculation, but was neither dose-dependent nor related to the extent of infection. Galactomannan was present in the plasma of infected mice and, in contrast to the situation in bovine placental aspergillosis, can be used as a good marker of foeto-placental aspergillosis.

Topics: Animals; Aspergillosis; Complement System Proteins; Female; Fetal Diseases; Galactose; Immunohistochemistry; Mannans; Mice; Mice, Inbred BALB C; Placenta Diseases; Pregnancy

The performance of the Pastorex Aspergillus antigen latex agglutination test for the detection of galactomannan in sera of patients at risk for invasive aspergillosis was evaluated, and the impact of storage on the reproducibility of the antigen titre was tested.. During a one year period, 392 serum samples were obtained from 46 patients at risk for invasive aspergillosis and tested for the presence of galactomannan using an Aspergillus latex agglutination test (Pastorex). Twenty three positive serum samples which had been stored at -20 degrees C for 2-16 months were retrospectively retested. Furthermore, two positive serum samples were stored at -20 degrees C and -70 degrees C and prospectively tested at three month intervals for a period of 15 months.. The Pastorex Aspergillus test was positive in eight patients with microbiological, radiological, or histological evidence for invasive aspergillosis, but was negative in the initial serum sample from five of these patients. In two patients with histological evidence for invasive aspergillosis no positive reaction was found in six samples. Six of 13 (45%) serum samples which had been stored at -20 degrees C for longer than six months had lost reactivity, while one of 10 (10%) samples had lost reactivity when stored up to six months. Two serum samples which had been stored at -20 degrees C and -70 degrees C and prospectively retested at three month intervals for 15 months, maintained stable antigen titres.. The Pastorex Aspergillus test is too insensitive to diagnose invasive aspergillosis in an early stage, but may contribute to the diagnosis when cultures remain negative and serial samples are obtained. To maintain a good reproducibility, serum samples should be stored at -70 degrees C when the period of storage exceeds six months.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Blood Preservation; Cryopreservation; Evaluation Studies as Topic; Female; Galactose; Humans; Latex Fixation Tests; Male; Mannans; Middle Aged; Prospective Studies; Reproducibility of Results; Temperature

A double-direct sandwich enzyme-linked immunosorbent assay that uses a rat anti-galactomannan monoclonal antibody as the acceptor and detector antibody was designed. This immunoassay, which detects less than 1 ng of galactomannan per ml, was assessed in a retrospective study with samples from patients with invasive aspergillosis. Serum is more appropriate than urine for use in the search for circulating galactomannan. Antigenemia does not have a transient character. Galactomannan can be detected at least 39 days before the death of the patients.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Child, Preschool; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; False Positive Reactions; Female; Galactose; Humans; Male; Mannans; Middle Aged; Opportunistic Infections; Sensitivity and Specificity

The G test containing factor G, fractioned from the Limulus lysate, was used to detect (1-3)-beta-D-glucan in a rat model of aspergillosis. Aspergillus fumigatus strain MF-13, 1 x 10(4) conidia, were inoculated transtracheally into rats treated with cortisone acetate (100 mg/kg) and fed a low-protein (8%) diet. Increased serum (1-3)-beta-D-glucan was found on the sixth day after inoculation in concentrations of 370 +/- 178 pg/ml (mean +/- SD) in untreated controls, and 154 +/- 43 pg/ml in rats treated with 0.5 mg/kg of amphotericin B. On day 11 (1-3)-beta-D-glucan concentrations were 2,590 +/- 2,940 pg/ml and 448 +/- 442 pg/ml, respectively. The elevation in levels of (1-3)-beta-D-glucan increased in correlation with the elevation of galactomannan antigen titers; (1-3)-beta-D-glucan is thus measurable during experimental aspergillosis in rats.

Topics: Amphotericin B; Animals; Aspergillosis; Aspergillus fumigatus; beta-Glucans; Disease Models, Animal; Galactose; Glucans; Horseshoe Crabs; Lung; Male; Mannans; Rats; Rats, Sprague-Dawley

The performance of a sandwich enzyme-linked immunosorbent assay (ELISA) which detects Aspergillus galactomannan (GM) was evaluated in bronchoalveolar lavage (BAL) fluid samples from 19 patients who were treated for hematological malignancies and who were suspected of having invasive pulmonary aspergillosis (IPA). All patients had fever and pulmonary infiltrates on the chest roentgenogram on the day that the BAL fluid was obtained. The ELISA results were compared with the results of culture and Aspergillus genus-specific PCR analysis of BAL fluid samples. ELISA was also performed with serum samples. Aspergillus species were detected by PCR or ELISA with BAL fluid samples from five of seven patients who had radiological evidence of IPA. Serum ELISA results were positive for all patients with ELISA-positive BAL fluid, and for four patients the serum ELISA was positive before the BAL fluid was obtained. PCR and ELISA were positive for 2 and 1 of 10 BAL fluid samples, respectively, obtained from patients without radiological evidence of IPA, and 5 and 2 of 35 BAL fluid samples, respectively, obtained from nonneutropenic patients. This preliminary investigation suggests that GM may be detected by ELISA in BAL fluid samples from patients at risk of IPA, but that monitoring of serum GM levels may allow for the earlier diagnosis of IPA. However, further evaluation in prospective studies is required.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; Aspergillus; Bronchoalveolar Lavage Fluid; Enzyme-Linked Immunosorbent Assay; Evaluation Studies as Topic; Female; Galactose; Hematologic Diseases; Humans; Leukemia; Lung Diseases, Fungal; Lymphoma; Male; Mannans; Middle Aged; Polymerase Chain Reaction; Sensitivity and Specificity

The detection of galactomannan antigen in urine was investigated in 26 bone marrow transplant recipients using an Aspergillus latex agglutination test (Pastorex). After modification of the method, which was originally devised for serum testing, the detection limit in native urine was approximately 20 ng/ml. Antigen was found in 79 (36.4%) of 217 serial urine samples, compared to 40 (11.8%) of 340 serum samples. As a rule, antigenuria preceded antigenemia and was more persistent. The sensitivity, specificity, positive predictive value and negative predictive value of antigenuria for autopsy-proven aspergillosis and clinically suspected Aspergillus infection were 57%, 53%, 31% and 77%, respectively, while those of antigenemia were 43%, 53%, 25% and 71%. It is concluded that urine testing is more reliable than serum testing for the detection of Aspergillus galactomannan. The detection of antigen, however, whether in serum or in urine, allows no clear distinction between Aspergillus infection and exposure to non-infectious Aspergillus antigens.

Topics: Adolescent; Adult; Antigens, Fungal; Aspergillosis; Aspergillus; Bone Marrow Transplantation; Female; Galactose; Humans; Male; Mannans; Middle Aged; Retrospective Studies

A commercial latex agglutination (LA) test for the detection of circulating Aspergillus galactomannan was evaluated in sera obtained from 121 immunocompromised patients, including 19 with proven invasive pulmonary aspergillosis. Patient urine (the specimen of choice for detection of galactomannan) was not tested with the LA test as 34 of 81 specimens from controls gave false-positive results. The sensitivity (95%), specificity (90%) and negative predictive value (99%) of the LA test were similar to previously published results obtained with two EIAs. However, the positive predictive value was only 67% compared to > or = 95% obtained with the EIAs. In addition, the LA test was also of less value than the EIAs in predicting the onset of invasive pulmonary aspergillosis. It was the earliest indicator of infection in only 1 of 19 proven cases.

Topics: Antigens, Fungal; Aspergillosis; Galactose; Humans; Immunocompromised Host; Immunoenzyme Techniques; Latex Fixation Tests; Lung Diseases, Fungal; Mannans; Retrospective Studies; Sensitivity and Specificity

The galactomannan (GM) produced extracellularly by Aspergillus fumigatus has been purified by a double sequential hydrazine-nitrous acid treatment of the ethanol precipitate of the culture filtrate. Nuclear magnetic resonance and gas-liquid chromatography-mass spectrometry analysis have been performed on intact GM, acid-hydrolyzed GM, and oligomers resulting from the acetolysis of the acid-hydrolyzed GM. Results show that A. fumigatus GM is composed of a linear mannan core with an alpha-(1-2)-linked mannotetraose repeating unit attached via alpha-(1-6) linkage. Side chains composed of an average of 4 to 5 beta-(1-5)-galactofuranose units are linked to C-6 and C-3 positions of alpha-(1-2)-linked mannose units of the mannan. The immunoreactivity of GM and HCl-hydrolyzed GM was studied by use of human sera from aspergillosis patients and an antigalactofuran monoclonal antibody. The alpha-(1-2) (1-6)-mannan core is not antigenic. The immunogenic galactofuran is found amongst several exocellular glycoproteins. According to a direct enzyme-linked immunosorbent assay with GM as the detector antigen, only 26% of the serum samples from aspergilloma patients (all positive by immunodiffusion assays) give optical density values superior to a cutoff estimated as the mean +/- 3 standard deviations of values obtained with control sera.

Topics: Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Carbohydrate Sequence; Chromatography, Gas; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Hydrolysis; Magnetic Resonance Spectroscopy; Mannans; Molecular Sequence Data

A model of primary pulmonary aspergillosis in rabbits was developed to reproduce the persistent levels of profound granulocytopenia and the histopathologic features of bronchopneumonia, vascular invasion, and hemorrhagic infarction encountered in humans. D-mannitol was detectable in bronchoalveolar lavage fluid by gas-liquid chromatography/mass spectroscopy, and galactomannan was measurable in serum by latex agglutination immunoassay. A pharmacokinetically distinctive unilamellar vesicle formulation of liposomal amphotericin B, 5 mg/kg/day intravenously, compared with high-dose conventional desoxycholate amphotericin B, 1 mg/kg/day intravenously, was more effective in preventing nephrotoxicity, increasing survival, reducing the number of viable organisms, and decreasing tissue injury due to Aspergillus organisms. Thus, D-mannitol in lavage fluid and galactomannan in serum may be useful markers of pulmonary aspergillosis, and liposomal amphotericin B was significantly more effective and safer than desoxycholate amphotericin B for treatment of pulmonary aspergillosis in profoundly granulocytopenic rabbits.

Topics: Agranulocytosis; Amphotericin B; Animals; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Biomarkers; Bronchoalveolar Lavage Fluid; Ergosterol; Galactose; Kidney Diseases; Life Tables; Liposomes; Lung Diseases; Mannans; Mannitol; Opportunistic Infections; Rabbits; Survival Analysis

(1-3)-beta-D-Glucan (beta-glucan) is a major structural component of fungi. The G test is a direct method to detect beta-glucan using fractionated (1-3)-beta-D-glucan-sensitive component, factor G, eluted from the limulus lysate. Previously, we reported that the G test is a more sensitive method than the mannan detection assay for the serological diagnosis of Candida infection. In this study, we discuss beta-glucanemia in patients with pulmonary aspergillosis and cryptococcosis. The concentration of beta-glucan was less than 10 pg/ml in 9 of 10 cases of pulmonary cryptococcosis, except for one case receiving hemodialysis (16.5 pg/ml). beta-Glucan increased in 3 cases of invasive pulmonary aspergillosis (27-937 pg/ml). Galactomannan antigen was positive in all of those cases. In 8 cases of aspergilloma, which showed fungus ball on roentgenogram, the mean concentration of beta-glucan was 67.1 +/- 92.7 pg/ml. Two of 8 cases were positive for galactomannan antigen. One of three cases of PAIC (productive aspergilloma on the inner wall of a cavity) and one case of chronic necrotizing pulmonary aspergillosis showed slightly increased levels of beta-glucan and positive results of galactomannan antigen test.

Topics: Adult; Aged; Antigens, Fungal; Aspergillosis; beta-Glucans; Child; Cryptococcosis; Female; Galactose; Glucans; Humans; Lung Diseases, Fungal; Male; Mannans; Middle Aged

Serum and urine samples from cattle with experimental and spontaneous systemic mycotic infections were tested for the presence of galactomannan and the 18 kDa antigen from Aspergillus fumigatus by an inhibition ELISA and immunoblotting, respectively. High levels of galactomannan (approximately 80 ng/ml.) were detected in serum from two of three calves experimentally infected with A. fumigatus. In two out of three cows with spontaneous acquired aspergillosis a similar amount of galactomannan was detected. Galactomannan was not found in serum samples of 20 cows which aborted due to either experimental or spontaneous placental aspergillosis. Twenty-four of forty urine samples from normal cattle reacted positively in the ELISA, consequently, the assay was not applicable on bovine urine. The 18 kDa antigen from A. fumigatus was detected in the urine from one calf out of three calves experimentally infected with aspergillosis and in the urine from one cow with spontaneous aspergillosis. It is concluded that detection of galactomannan in serum and the 18 kDa antigen in urine may be used as an aid for the diagnosis of disseminated bovine aspergillosis with the exception of placental and probably gastrointestinal localization.

Topics: Abortion, Veterinary; Animals; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Cattle; Cattle Diseases; Female; Galactose; Male; Mannans; Pregnancy; Pregnancy Complications, Infectious

To improve the immunohistopathological diagnosis of systemic bovine mycoses, we have evaluated the utility of antifungal polyclonal and monoclonal antibodies, and peroxidase and alkaline phosphatase staining techniques. A rabbit polyclonal antibody to mannan from Candida albicans was specific for candidosis. The diagnosis of aspergillosis was accomplished using a rat monoclonal antibody to the galactofuran side chains of Aspergillus galactomannan. A murine monoclonal antibody reacting with weakly Con-A binding 41 and 46 kDa somatic antigens from Absidia corymbifera was used for immunostaining of zygomycetic hyphae. Peroxidase antiperoxidase (PAP) and alkaline phosphatase antialkaline phosphatase (APAAP) complexes were visualized using aminoethylcarbazole and fast red substrates. A green staining of PAP reactions with dioctyl sulfosuccinate sodium and 3,3',5,5'-tetramethylbenzidine (DONS/TMB) was effective for the demonstration of fungi in dual and triple infections. Tissue sections of experimentally infected mice were used to determine the sensitivity and specificity of the antibodies. Tissues obtained from 161 bovine mycotic lesions previously studied by indirect immunofluorescence staining were further evaluated using the three antibodies. In all of 45 lesions solely affected by aspergillosis and in three solely affected by candidosis the diagnoses were confirmed by the new evaluation. In 85 of 96 cases of single infections with zygomycetes the diagnosis was confirmed, while none of the antibodies reacted with fungal elements in the remaining 11 lesions. Aspergillus hyphae were detected in all three lesions with dual aspergillosis and zygomycosis, whereas zygomycetic material was confirmed in only two of these cases. A mixed infection of candidosis and zygomycosis in a lymph node was confirmed too. In 13 cases in which a diagnosis had not hitherto been obtained, aspergillosis and zygomycosis were recorded each in three cases.

Topics: Animals; Antibodies; Antibodies, Monoclonal; Antigens, Fungal; Aspergillosis; Aspergillus; Candidiasis; Cattle; Cattle Diseases; Female; Galactose; Immunoenzyme Techniques; Mannans; Mice; Mice, Inbred BALB C; Mycoses; Omasum; Placenta; Pregnancy; Rabbits

The monoclonal antibody EB-A1 to galactomannan is apparently specific for detecting Aspergillus species and Penicillium marneffei in formalin-fixed, paraffin-embedded tissues. It reveals hyphae, remnants of filaments, and organisms in the cytoplasm of some phagocytic cells.

Topics: Antibodies, Monoclonal; Aspergillosis; Cell Wall; Cytoplasm; Dermatomycoses; Galactose; Humans; Immunohistochemistry; Liver Diseases; Lung Diseases, Fungal; Mannans; Phagocytes

The performance of Pastorex Aspergillus, a new latex agglutination test for the detection of circulating galactomannan in the serum of patients with invasive aspergillosis, was evaluated in a blind trial in standardized guinea-pig models of invasive aspergillosis and other invasive mycoses. In these animal models, the invasive nature of the fungal infection was confirmed by re-isolation of the etiologic agent from the organs of every animal. Ninety-two plasma samples from 42 animals with invasive aspergillosis were submitted to the test. In 41 of these animals, at least one plasma sample was positive with the latex test (sensitivity 97.6%), titers ranging from 1/1 to 1/512. In general, antigen titers increased as a function of time, reaching the highest values shortly before death. Guinea-pigs infected with Penicillium marneffei also yielded positive agglutination reactions but antigen titers were lower (maximal titer 1/8). Plasma samples from animals with invasive candidosis (23), disseminated trichophytosis (11) and cryptococcosis (23) were all negative with the latex test. In 80 guinea-pigs without fungal infection, 3 false positive results (titers 1/1) were observed, which means a specificity of 96.2% in this control group.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Disease Models, Animal; Galactose; Guinea Pigs; Latex Fixation Tests; Male; Mannans; Reagent Kits, Diagnostic

Purified galactomannan (GM) from Aspergillus fumigatus was used in both a radioimmunoassay and an enzyme-linked immunoassay for antigen detection. Results of the two tests seemed interchangeable. By one or both assays, GM was detected in serum from four of 12 rabbits lethally infected with A. fumigatus in concentrations ranging from 108 to 356 ng/ml. Serum antigen was detected in only two of 12 patients with invasive aspergillosis. Results of assay for GM in urine were far more encouraging. Urinary GM was detectable throughout the course of lethal aspergillosis in all 16 rabbits, in concentrations of 24-1,900 ng/ml. Urine from seven of 13 patients with invasive aspergillosis had GM concentrations of 1-83 ng/ml. Antigen excretion roughly paralleled extent of disease.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus flavus; Aspergillus fumigatus; Enzyme-Linked Immunosorbent Assay; Galactose; Humans; Mannans; Rabbits; Radioimmunoassay

A sensitive enzyme immunoassay (EIA) for galactomannan antigenemia that avoids the use of radioisotopes was devised. Three carbohydrate-rich antigenic fractions were purified from Aspergillus fumigatus 2085: a cold alkali extract (CA) from mycelium, an acetone-precipitated pyridine extract (APSK-66) from mycelium, and a methanol precipitate from culture filtrate. CA and APSK-66 were further purified by gel filtration and ion-exchange chromatography, respectively. An acid hydrolysate of CA contained only mannose and galactose, as determined by gas-liquid chromatography. Rabbit antisera were raised against conidia, mycelia, and cell walls of A. fumigatus. By indirect EIA, the best immunoglobulin G response (1/8,000) was obtained against CA in rabbits immunized intravenously with cell walls. Antigenemia was detected by indirect EIA inhibition in heat-dissociated sera of four immunosuppressed rabbits that were infected intravenously but was absent in two uninfected controls. The circulating antigen was resistant to pronase, was adsorbed onto concanavalin A, and had a molecular size of 50 to 100 kilodaltons.

Topics: Animals; Antibodies, Fungal; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Chromatography, Affinity; Chromatography, Gas; Chromatography, Gel; Chromatography, Ion Exchange; Female; Galactose; Immunoenzyme Techniques; Immunoglobulin G; Immunosuppression Therapy; Mannans; Rabbits; Ultrafiltration

Galactomannan (GM) extracted from mycelia of Aspergillus fumigatus with cold dilute alkali reacted with antiserum specific for an antigen that circulated in invasive aspergillosis in rabbits and humans. The GM was purified by its affinity for concanavalin A and was separated from a nonantigenic glucan by gel permeation on Sephacryl S-200. The GM molecular weight of between 25,000 to 75,000 was smaller than the antigen present in infected rabbit serum which was retained by an ultrafiltration membrane that had a nominal molecular weight limit of 125,000. The ratio of galactose to mannose present in GM was 1:1.17. The serological activity of GM was stable to boiling but labile to 0.01 N HCl, implicating galactofuranose as an antigenic determinant. Analysis of purified GM by methylation-gas chromatography suggested a structure consisting of a 1 leads to 6-linked mannan backbone with oligogalactoside side chains 3 units long, terminating in galactofuranose. The presence of mannose as a side chain component was also inferred. Another antigen of A. fumigatus, which did not bind to concanavalin A, was isolated after tandem chromatography on diethylaminoethyl- and carboxymethyl-Sephadex and was identified as a galactan. The galactan inhibited the immune precipitation of GM was specific antiserum.

Topics: Animals; Antigens, Fungal; Aspergillosis; Aspergillus fumigatus; Chemical Phenomena; Chemistry; Chromatography, Affinity; Galactans; Galactose; Humans; Mannans; Methylation; Molecular Weight; Monosaccharides; Polysaccharides; Rabbits; Ultrafiltration