leukotoxin and Dental-Plaque

Mechanical debridement results in a shift of the bacterial composition in the periodontal pocket on the species level. It is unknown, however, whether a clonal change within a species could lead to the emergence of strains with different levels of virulence. Therefore, in the present study, the genetic variability of Actinobacillus actinomycetemcomitans was assessed and strains identified which were associated with periodontal disease progression following periodontal therapy, i.e., refractory periodontitis. Twenty adult patients with untreated periodontitis and subgingival colonization of A. actinomycetemcomitans were randomly assigned to receive full-mouth scaling alone or scaling with an adjunctive antimicrobial therapy. Both groups received supportive periodontal therapy at 3, 6, 9, 12, 18, and 24 months. Subgingival plaque samples were taken at every visit; venous blood was obtained at 24 months only. A. actinomycetemcomitans isolates were typed by the RAPD method, and antibody reactivity against outer membrane proteins was assessed by immunoblot analysis. Eleven distinct RAPD patterns were found in 18 patients completing the study. All patients harbored only one A. actinomycetemcomitans genotype, and within each patient this genotype persisted throughout the 24-month observation period. No differences in the expression of antibody reactivity against outer membrane proteins were found between strains isolated at baseline and at 24 months. Three genotypes were associated with reduced survival rates of teeth without probing attachment loss of 2 mm or more. The results indicated that (i) most patients harbored only one A. actinomycetemcomitans genotype; (ii) the genotype persisted following therapy; and (iii) only some genotypes were associated with refractory periodontitis.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Amoxicillin; Anti-Bacterial Agents; Bacterial Outer Membrane Proteins; Bacterial Typing Techniques; Chronic Disease; Clone Cells; Dental Plaque; Disease Progression; DNA Fingerprinting; DNA, Bacterial; Exotoxins; Female; Genetic Variation; Humans; Male; Metronidazole; Middle Aged; Penicillins; Periodontal Pocket; Periodontitis; Random Amplified Polymorphic DNA Technique; Survival Analysis; Virulence

To assess the progression of attachment loss (AL) during a 2-year period according to the presence of JP2 and non-JP2 genotypes of Aggregatibacter actinomycetemcomitans in a Ghanaian adolescent population.. A total of 500 adolescents (mean age 13.2 years, SD ± 1.5) were enrolled in the study. After 2 years, 397 (79.4%) returned for a periodontal re-examination, including the measurement of AL. The carrier status of the JP2 and non-JP2 genotypes of A. actinomycetemcomitans was evaluated in a baseline examination 2 years earlier.. Individuals who carried the JP2 genotype of A. actinomycetemcomitans had a significantly increased risk [relative risk (RR) = 7.3] of developing AL ≥ 3 mm. The mean AL at the follow-up and the mean 2-year progression of AL were significantly higher in the JP2 genotype-positive group (n = 38) compared with the group positive for the non-JP2 genotypes of A. actinomycetemcomitans (n = 169), and the group of A. actinomycetemcomitans-negative individuals (n = 190). The JP2 genotype was strongly associated with the progression of AL ≥ 3 mm (OR = 14.3). The non-JP2 genotypes of A. actinomycetemcomitans were also, however, less pronounced, associated with the progression of AL ≥ 3 mm (OR = 3.4).. The JP2 genotype of A. actinomycetemcomitans is strongly associated with the progression of AL.

Topics: Adolescent; Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Cohort Studies; Cytotoxins; Dental Plaque; Disease Progression; Exotoxins; Female; Follow-Up Studies; Genotype; Ghana; Humans; Male; Periodontal Attachment Loss; Prospective Studies; Risk Factors; Virulence Factors

The aim of this study was to characterize the main periodontal bacterial species in Down syndrome (DS) patients with and without periodontitis.. This cross-sectional study involved 75 DS patients, 45 with and 30 without periodontitis. Informed consent, health and dental questionnaires and periodontitis diagnosis were performed PCR and LAMP assays were performed on subgingival dental plaque sample.. Tannerella forsythia was the most frequent bacteria detected in the group with and without periodontitis (95.5 and 63.3%) followed by Treponema denticola (88.8 and 50%) and Porphyromonas gingivalis (53.3 and 25% respectively). There were statistical differences between groups (p < 0.05). Pg fimA type I was the most frequent Porphyromonas gingivalis genotype. Two different sets of primers (Aa-F/Aa-R and ltx3/ltx4) were used to detect Aggregatibacter actinomycetemcomitans and different frequencies were obtained, (68% and 14.6% respectively), they had a weak correlation (Cohen Kappa = 0.16). After sequencing of PCR products, ltx3/ltx4 showed more specificity. JP2 clone of A. actinomycetemcomitans was not detected in any sample.. The composition of oral biofilm is fundamental for the development of periodontal disease independently of immunological alterations associated with DS. The frequency of detection of A. actinomycetemcomitans reported in the literature has a wide range, because the primers and probes applied

Topics: Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Bacteroides; Biofilms; Cross-Sectional Studies; Dental Plaque; DNA Primers; DNA, Bacterial; Down Syndrome; Exotoxins; Female; Fimbriae Proteins; Genotype; Humans; Male; Microbial Consortia; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Periodontium; Pili, Sex; Porphyromonas gingivalis; Tooth Loss; Treponema denticola; Young Adult

To investigate the presence of A. actinomycetemcomitans, including the highly virulent JP2 clone, in young adult patients with aggressive periodontitis, and associate the findings with the two forms of the disease.. Seventy Moroccan subjects with aggressive periodontitis, aged less than 35 years, were recruited. Among these, 41 had LAgP and 29 had GAgP. Plaque samples were collected from periodontal pockets and examined using a PCR that detects the presence of A. actinomycetemcomitans and which differentiates between JP2 and non-JP2 genotypes of the bacterium.. total of 58 (83%) from the 70 AgP patients were positive for A. actinomycetemcomitans, among whom 77% were positives for the JP2 clone. The JP2 clone was detected in 34 (83%) of the LAgP patients compared to 20 (69%) of the GAgP patients (p = 0.17). Fourteen (20%) of the patients harbored non-JP2 genotypes of A. actinomycetemcomitans, although most of these patients (10/14) also harbored the JP2 clone.. The presence of the JP2 clone of A. actinomycetemcomitans is strongly associated with both LAgP and GAgP in young adults in Morocco. This implies that treatment of AgP in this population should include microbiological screening and aim at eradication of the bacterium when present.

Topics: Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Bacterial Toxins; Chi-Square Distribution; Child; Dental Plaque; Exotoxins; Female; Genotype; Humans; Male; Morocco; Periodontal Pocket; Promoter Regions, Genetic; Virulence; Young Adult

Limited data are reported concerning the presence of A. actinomycetemcomitans and attachment loss (AL) in sub-Saharan countries. The authors investigate the carrier frequency of JP2 and non-JP2 genotypes of A. actinomycetemcomitans and the presence of AL in Ghanaian adolescents and evaluate socioeconomic conditions and oral hygiene practices.. Five hundred individuals (mean ± SD age: 13.2 ± 1.5 years) in public and private schools were interviewed about demographic characteristics and oral hygiene practices and were given a full-mouth periodontal examination. Subgingival plaque samples were obtained from periodontal sites around permanent first molars and incisors. The carrier status of A. actinomycetemcomitans at the individual level was determined based on results obtained by cultivation and polymerase chain reaction.. The findings of this study show a relatively high carrier rate of JP2 and non-JP2 genotypes of Aggregatibacter actinomycetemcomitans in the Ghanaian adolescent population and the presence of this bacterium is associated with the occurrence of AL. The overall carrier rate of A. actinomycetemcomitans was 54.4%, and the highly leukotoxic JP2 genotype was detected in 8.8% of the study population. A total of 107 (21.4%) individuals had ≥ 1 tooth with AL ≥ 3 mm. The majority of the individuals carrying A. actinomycetemcomitans (80.1%) (P <0.001) and of the periodontally diseased individuals (91.6%) (P <0.001) were found in public schools.. A. actinomycetemcomitans and AL were frequently found in Ghanaian adolescents. The school type was the strongest predictor of both presence of A. actinomycetemcomitans and AL.

Topics: Adolescent; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Carrier State; Child; Confidence Intervals; Cross-Sectional Studies; Dental Plaque; Exotoxins; Female; Genotype; Ghana; Humans; Logistic Models; Male; Oral Hygiene; Periodontal Attachment Loss; Private Sector; Public Sector; Schools; Sequence Deletion; Social Class; Surveys and Questionnaires; Young Adult

To examine the genetic diversity of Aggregatibacter actinomycetemcomitans in Thai adults.. Subgingival plaque samples from 453 subjects were analysed for A. actinomycetemcomitans serotypes, the presence of the high leukotoxin-producing JP2 clone and cytolethal distending toxin genes (cdtABC) using the polymerase chain reaction technique. In subjects who were positive for cdtABC, restriction fragment length polymorphism analysis was used to identify a single nucleotide polymorphism (SNP) in the cdtB gene at amino acid position 281. The extent and severity of periodontal disease were compared between subjects harbouring different A. actinomycetemcomitans genotypes.. Eighty six subjects (19%) were positive for A. actinomycetemcomitans. The JP2 clone was not detected. Serotype c was the most prevalent (57%), followed by serotypes a (33%) and b (7%). Among A. actinomycetemcomitans-positive subjects, 27% were positive for cdtABC. All cdtABC-positive subjects possessed the SNP in the cdtB, which is involved with increased toxin activity. The presence of A. actinomycetemcomitans, but not a specific genotype, was significantly related to increased probing depth and periodontal attachment loss.. Our results confirm the previous findings that genotype distribution of A. actinomycetemcomitans varies between ethnic groups. However, no clear relationship between a specific genotype and periodontal conditions was observed.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Analysis of Variance; Bacterial Toxins; Chi-Square Distribution; Cross-Sectional Studies; Dental Plaque; Exotoxins; Female; Humans; Male; Middle Aged; Molecular Epidemiology; Periodontal Attachment Loss; Periodontitis; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide; Serotyping; Thailand

carriers of the JP2 clone of Aggregatibacter actinomycetemcomitans exhibit an enhanced risk for developing aggressive periodontitis compared with individuals carrying non-JP2 clones. While the JP2 clone is almost exclusively detected among adolescents of African descent, reports on Caucasians colonized with the JP2 clone are remarkably few.. the aim of this paper is to report on the history of periodontal disease and microbiological findings in a Caucasian family.. a. actinomycetemcomitans and other periodontitis-associated bacterial species in subgingival plaque samples were quantified by conventional culture technique. Leucotoxin promoter typing, serotyping and further characterizations of A. actinomycetemcomitans isolates were performed by PCR. DNA sequencing of the pseudogene, hbpA was performed to determine the origin of the detected JP2 clones. Further, genetically ancestry testing of family members was carried out.. the JP2 clone was detected in samples from two of the family members, a 33-year-old daughter and her 62-year-old mother. Relationship of their JP2 clones with JP2 clone strains from the Mediterranean area of Africa was indicated. Genotyping confirmed the Caucasian origin of all family members.. Caucasian JP2 carriers exist and older subjects can carry the JP2 clone of A. actinomycetemcomitans.

Topics: Actinobacillus Infections; Adult; Aggregatibacter actinomycetemcomitans; Chronic Periodontitis; Dental Plaque; Exotoxins; Female; Follow-Up Studies; Humans; Middle Aged; Serotyping; Subgingival Curettage; White People

Aggregatibacter actinomycetemcomitans is considered a possible etiological agent for aggressive periodontitis. The aim of this study was to determine the prevalence of the JP2 clone and non-JP2 genotypes of A. actinomycetemcomitans in the subgingival plaque of patients with aggressive periodontitis and controls among Sudanese high-school students.. In a previous study we examined a large representative sample of students attending high schools in Khartoum, Sudan. In this population, 17 patients with aggressive periodontitis and 17 controls (14-19 years of age) consented to participate in the present study. The subjects underwent a clinical periodontal examination, and subgingival dental plaque samples were collected using paper points. The presence of the A. actinomycetemcomitans JP2 clone and non-JP2 genotypes were assessed using loop-mediated isothermal amplification (LAMP) and the PCR.. The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of either the cases or the controls. Non-JP2 types of A. actinomycetemcomitans were detected in the subgingival plaque of 12 (70.6%) patients with aggressive periodontitis and from only one (5.9%) control subject, showing a significantly higher frequency of detection in cases than in controls (p = 0.0001). The odds ratio for the detection of A. actinomycetemcomitans in the subgingival plaque of the patients with aggressive periodontitis was 38.4 (95% confidence interval: 4.0-373.0; p = 0.002). The PCR and LAMP methods showed identical results pertaining to the identification of non-JP2 types of A. actinomycetemcomitans.. The JP2 clone of A. actinomycetemcomitans was not detected in the subgingival plaque of high school subjects in Sudan. The detection of non-JP2 types of A. actinomycetemcomitans may be a useful marker of increased risk for development of aggressive periodontitis in young subjects.

Topics: Actinobacillus Infections; Adolescent; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Bacterial Toxins; Case-Control Studies; Clone Cells; Dental Plaque; DNA, Bacterial; Exotoxins; Female; Genotype; Humans; Male; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; RNA, Bacterial; RNA, Ribosomal, 16S; Sudan; Young Adult

The influence of Aggregatibacter actinomycetemcomitans on inflammation in subjects with gingivitis has not been studied in great detail. Seventeen healthy young adults with plaque-induced gingivitis or localized mild chronic periodontitis harboring cultivable numbers of A. actinomycetemcomitans were thoroughly examined. Samples of subgingival plaque were obtained from mesial surfaces of all teeth present. In addition, 12 oral mucosal surfaces and unstimulated saliva were sampled. Species identity, presence of the leukotoxin gene, and absence of a specific 530 b deletion in the leukotoxin promoter region indicating non-JP2-like strains were assessed by polymerase chain reaction. Based on a multilevel random intercept model adjusted for probing depth, age, and smoking status, the odds of bleeding on probing was increased by a factor of 1.89 (1.09-3.29, p = 0.024) if, in addition to plaque, A. actinomycetemcomitans could be recovered from the site. At a site without visible supragingival plaque but with cultivable numbers of subgingival A. actinomycetemcomitans the odds ratio of bleeding on probing was 3.37 (0.86-13.2, p = 0.081). Simulating variance partition coefficients revealed that between 1-2% (a clean, shallow site without A. actinomycetemcomitans; a deep site covered by plaque containing A. actinomycetemcomitans) and 6-7% (a moderately deep site with neither visible plaque nor cultivable A. actinomycetemcomitans) of the residual variance was attributable to differences between subjects. The present cross-sectional study indicates that non-JP2-like strains of A. actinomycetemcomitans may enhance gingival bleeding tendency even in the absence of clinically visible supragingival plaque.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Colony Count, Microbial; Cross-Sectional Studies; Dental Plaque; Exotoxins; Female; Gingivitis; Humans; Male; Multilevel Analysis; Periodontal Index; Promoter Regions, Genetic; Sequence Deletion; Species Specificity; Young Adult

To look for clinical signs of periodontal disease in young adults who exhibited radiographic bone loss and detectable numbers of Aggregatibacter actinomycetemcomitans in their primary dentition.. Periodontal status and radiographic bone loss were examined in each of the subjects 16 years after the baseline observations. Techniques for anaerobic and selective culture, and checkerboard, were used to detect periodontitis-associated bacterial species. The isolated A. actinomycetemcomitans strains were characterized by polymerase chain reaction.. Signs of localized attachment loss were found in three out of the 13 examined subjects. A. actinomycetemcomitans was recovered from six of these subjects and two of these samples were from sites with deepened probing depths and attachment loss. Among the isolated A. actinomycetemcomitans strains, serotypes a-c and e, but not d or f, were found. None of the isolated strains belonged to the highly leucotoxic JP2 clone, and one strain lacked genes for the cytolethal distending toxin.. This study indicates that the presence of A. actinomycetemcomitans and early bone loss in the primary dentition does not necessarily predispose the individual to periodontal attachment loss in the permanent dentition.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Alveolar Bone Loss; Bacterial Toxins; Bacteroides; Clone Cells; Colony Count, Microbial; Cytotoxins; Dental Calculus; Dental Plaque; DNA, Bacterial; Exotoxins; Female; Follow-Up Studies; Genotype; Humans; Male; Nucleic Acid Hybridization; Peptostreptococcus; Periodontal Attachment Loss; Periodontal Pocket; Porphyromonas gingivalis; Prevotella intermedia; Prevotella nigrescens; Serotyping; Young Adult

The identification of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans) serotypes may add some important information to the understanding of the pathogenetic background of severe periodontal infections. This study compared serotypes of A. actinomycetemcomitans in two groups of periodontal patients with different ethnic backgrounds.. A total of 194 patients (96 Germans and 98 Koreans) with aggressive or severe chronic periodontitis participated in the study. Microbiologic analysis of pooled samples from subgingival plaque was performed by using a real-time polymerase chain reaction (PCR) test for A. actinomycetemcomitans. In patients who tested positive for A. actinomycetemcomitans, serotypes (a through f) were determined by nucleic acid-based methods.. The prevalence of patients who tested positive for A. actinomycetemcomitans with the real-time PCR was comparable in both groups (Germans: 27.0%; Koreans: 22.2%). In German patients, the serotypes detected most frequently were b (33.3%), c (25.0%), and a (20.8%), whereas in Korean patients, the serotype distribution was different, with serotypes c (61.9%) and d (19.0%) accounting for >80% of the complete serotype spectrum.. Even if the percentage of patients who tested positive for A. actinomycetemcomitans was identical in patients with generalized aggressive and severe chronic periodontitis and different ethnic backgrounds, the distribution of A. actinomycetemcomitans serotypes may exhibit marked differences.

Topics: Actinobacillus Infections; Adult; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Asian People; Bacterial Toxins; Chronic Periodontitis; Dental Plaque; Dental Plaque Index; Ethnicity; Exotoxins; Female; Germany; Gingival Hemorrhage; Humans; Korea; Male; Middle Aged; Operon; Periodontal Attachment Loss; Periodontal Index; Periodontal Pocket; Polymerase Chain Reaction; Promoter Regions, Genetic; Serotyping; White People

beta2-Glycoprotein I (beta 2GPI) is important in the suppression of coagulation, and antibodies against TLRVYK peptides on the beta 2GPI molecule are related to thrombosis. According to the Swiss-Prot database, Aggregatibacter actinomycetemcomitans leukotoxin c has sequences (SIRVYK) that are homologous to the TLRVYK peptides. The aim of this study was to investigate the effects of A. actinomycetemcomitans infection on the antibody response against SIRVYK peptides in patients with periodontitis.. Serum immunoglobulin G (IgG) antibody and IgG subclass antibody titers against SIRVYK or TLRVYK peptides were measured by enzyme-linked immunosorbent assay in 46 patients with aggressive periodontitis (eight with localized disease, 38 with generalized disease), 28 patients with chronic periodontitis, and 20 periodontally healthy subjects. The presence of A. actinomycetemcomitans in plaque and saliva samples was determined using polymerase chain reaction.. The level of anti-SIRVYK antibodies was significantly higher in patients who were A. actinomycetemcomitans-positive than in A. actinomycetemcomitans-negative patients (P < 0.05) in the chronic periodontitis group. A similar trend was found in the antibody response to TLRVYK peptide; however, no statistically significant difference was seen between A. actinomycetemcomitans-positive and -negative patients. The A. actinomycetemcomitans-positive patients displayed significantly higher levels of anti-SIRVYK IgG2 and IgG3 antibodies than A. actinomycetemcomitans-negative patients (P < 0.05 and P < 0.05, respectively). The level of IgG2 was highest among the four IgG subclasses and it predominantly increased in patients who were A. actinomycetemcomitans-positive. Anti-TLRVYK antibody levels were significantly correlated with anti-SIRVYK IgG antibody levels.. The results suggest that A. actinomycetemcomitans infection may elicit anti-SIRVYK IgG antibodies and modify the anti-TLRVYK antibody response in patients with periodontitis by molecular mimicry with beta2GPI.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Antibodies, Bacterial; Anticoagulants; Bacterial Toxins; beta 2-Glycoprotein I; Chronic Disease; Dental Plaque; Exotoxins; Gingival Hemorrhage; Humans; Immunoglobulin G; Middle Aged; Molecular Mimicry; Peptide Fragments; Periodontal Pocket; Periodontitis; Periodontium; Polymerase Chain Reaction; Saliva

The aim of our study is to describe a fast molecular method, able to distinguish and quantize the two different genotypes (652 and JP2) of an important periodontal pathogen: Actinobacillus actinomycetemcomitans. The two genotypes show differences in the expression of an important pathogenic factor: the leukotoxin (ltx). In order to evidence this, we performed a real time PCR procedure on the ltx operon, able to recognize Aa clinical isolates with different leukotoxic potentials.. The specificity of the method was confirmed in subgingival plaque and saliva specimens collected from eighty-one Italian (Sardinian) subjects with a mean age of 43.9, fifty five (68 %) of whom had various clinical forms of periodontal disease.. This procedure showed a good sensitivity and a high linear dynamic range of quantization (10(7)-10(2) cells/ml) for all genotypes and a good correlation factor (R2 = 0.97-0.98). Compared with traditional cultural methods, this real time PCR procedure is more sensitive; in fact in two subgingival plaque and two positive saliva specimens Aa was only detected with the molecular method.. A low number of Sardinian patients was found positive for Aa infections in the oral cavity, (just 10 positive periodontal cases out of 81 and two of these were also saliva positive). The highly leukotoxic JP2 strain was the most representative (60 % of the positive specimens); the samples from periodontal pockets and from saliva showed some ltx genotype for the same patient. Our experience suggests that this approach is suitable for a rapid and complete laboratory diagnosis for Aa infection.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aggregatibacter actinomycetemcomitans; Benzothiazoles; Child; Dental Plaque; Diamines; Exotoxins; Female; Genotype; Humans; Male; Middle Aged; Organic Chemicals; Periodontal Diseases; Polymerase Chain Reaction; Quinolines; Saliva; Sensitivity and Specificity

The use of whole saliva has shown to be promising in detecting Actinobacillus actinomycetemcomitans out of the subgingival environment. The objective of the present study was to evaluate the use of unstimulated saliva in detecting A. actinomycetemcomitans and to compare the subgingival and extracrevicular occurrence of this pathogen in Brazilian subjects with chronic periodontitis.. Sixty-six patients (mean age 38.01 9.28 years) with advanced generalized chronic periodontitis were sampled. Subgingival plaque samples were collected from eight sites per patient representing the two deepest sites of each quadrant. Samples of the mucous surfaces, including dorsal surface of the tongue and cheek, were collected with a sterile swab and placed in a microtube containing a reduced solution. Samples of unstimulated saliva were also collected in sterile tubes and 0.1 ml of whole saliva was diluted in 1 ml of reduced solution. The presence of A. actionomycetemcomitans was established using bacterial culture in trypticase soy bacitracin vancomycin selective media. Polymerase chain reaction (PCR) was used to differentiate highly from minimally leukotoxic strains in patients who presented A. actinomycetemcomitans in at least two sampled sites.. A. actinomycetemcomitans was isolated from 63.63% of subgingival samples, 56.06% of saliva samples, and 45.45% of samples from mucous surfaces. No statistical difference was observed between subgingival and salivary occurrence of the microorganism. Linear regression showed an association between subgingival plaque and saliva (r(2) = 0.897; P = 0.015) and mucous membrane and saliva (r(2) = 0.152; P = 0.024). The same A. actinomycetemcomitans leukotoxic profile was observed in all sampled sites for a given patient.. These results suggest that in advanced periodontitis, unstimulated saliva is representative of pooled subgingival plaque samples and its use is appropriate in the oral detection of A. actinomycetemcomitans.

Topics: Adult; Aggregatibacter actinomycetemcomitans; Chronic Disease; Dental Plaque; Exotoxins; Female; Humans; Linear Models; Male; Middle Aged; Mouth Mucosa; Periodontitis; Polymerase Chain Reaction; Saliva

Previous studies suggest differences between geographically and racially distinct populations in the prevalence of periodontopathic bacteria as well as greater periodontal destruction associated with infection by highly leucotoxic Actinobacillus actinomycetemcomitans. The present study examined these hypotheses in Brazilians with aggressive or chronic periodontitis.. Clinical, radiographical, and microbiological assessments were performed on 25 aggressive periodontitis and 178 chronic periodontitis patients including 71 males and 132 females, 15-69 years of age.. The prevalence of Porphyromonas gingivalis was similar to that of other South American populations. The prevalence of A. actinomycetemcomitans and its highly leucotoxic subgroup was higher in Brazilians. Highly leucotoxic A. actinomycetemcomitans was more prevalent in aggressive periodontitis (chi2=27.83) and positively associated with deep pockets (>6 mm, chi2=18.26) and young age (<29 years, chi2=18.68). Greater mean attachment loss was found in subjects with highly leucotoxic A. actinomycetemcomitans than in subjects with minimally leucotoxic (p=0.0029) or subjects not infected (p=0.0001).. These data support the hypothesis of differences between populations in the prevalence of periodontopathic bacteria and of greater attachment loss in sites infected with highly leucotoxic A. actinomycetemcomitans. Detection of highly leucotoxic A. actinomycetemcomitans in children and adolescents may be a useful marker for aggressive periodontitis.

Topics: Acute Disease; Adolescent; Adult; Aged; Aggregatibacter actinomycetemcomitans; Bacteroides; Brazil; Campylobacter rectus; Chronic Disease; Colony Count, Microbial; Cross-Sectional Studies; Dental Plaque; DNA, Bacterial; Exotoxins; Female; Humans; Logistic Models; Male; Middle Aged; Periodontitis; Porphyromonas gingivalis; Prevalence; Prevotella intermedia

The JP2 clone of Actinobacillus actinomycetemcomitans, a high-leukotoxin-producing strain, characterized by a 530-basepair (bp) deletion in the promoter region of the leukotoxin gene operon and mainly found among individuals with African origin, is associated with localized aggressive periodontitis. The objective of the study was to examine the occurrence of periodontal disease in a Moroccan immigrant family living in Denmark in which the oldest son (14 year) was referred and treated for localized aggressive periodontitis. Further, the potential occurrence of the JP2 clone of A. actinomycetemcomitans in the family was examined. Here we present the clinical, radiographic, and microbiological findings from the family. Clinical and radiographic examination of the other family members revealed that 3 of 5 younger siblings had localized aggressive periodontitis, one had gingivitis and the mother had chronic periodontitis. Despite scaling followed by intensive maintenance therapy several family members, including the sibling with gingivitis, had further attachment loss at the 1-year examination. The JP2 clone of A. actinomycetemcomitans was isolated from subgingival plaque samples from 4 children with periodontitis. In contrast, it was not detected in plaque from the oldest boy, who had been treated for localized aggressive periodontitis by surgery combined with antibiotic therapy. The 4 children with periodontitis and colonized with the JP2 clone were treated by scaling and antibiotic administration. One month later the JP2 clone could still be detected in plaque samples. In conclusion, it is confirmed that members of immigrant families with African origin are potential carriers of the JP2 clone and that those families often have multiple family members with localized aggressive periodontitis. It is proposed that those families are given periodontal examination frequently to benefit from early diagnosis and treatment of the disease.

Topics: Actinobacillus Infections; Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Base Sequence; Child; Chronic Disease; Denmark; Dental Plaque; Emigration and Immigration; Exotoxins; Female; Follow-Up Studies; Gingivitis; Humans; Male; Morocco; Operon; Periodontitis; Promoter Regions, Genetic; Sequence Deletion

The JP2 clone of Actinobacillus actinomycetemcomitans is associated with early-onset periodontitis in certain ethnic populations of African origin. Here, we describe and evaluate a set of primers for PCR to assay for the presence of A. actinomycetemcomitans and to discriminate between JP2-like strains and other genotypes in subgingival plaque samples.

Topics: Actinobacillus Infections; Adolescent; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Dental Plaque; DNA Primers; Exotoxins; Gingiva; Humans; Polymerase Chain Reaction; Sensitivity and Specificity

A clone of Actinobacillus actinomycetemcomitans (JP2) with increased leukotoxin production and characterized by a 530-bp deletion in the leukotoxin gene operon is endemically present in Morocco and strongly associated with the presence of early onset periodontitis (EOP).. To compare patterns of attachment loss among EOP-patients with or without JP2-type of A. actinomycetemcomitans in dental plaque.. Among 45 Moroccan adolescents with EOP (i.e. one or more teeth with attachment loss >/= 3 mm) 39 had cultivable plaque samples. Fifteen (38.5%) were culture-positive for A. actinomycetemcomitans of the JP2-type as determined by PCR, and 24 (61.5%) were not (mean age 16.5 years in both groups).. EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type had significantly more teeth with attachment loss (mean 5.1, median 4.0) than EOP-patients not culture-positive for A. actinomycetemcomitans of the JP2-type (mean 2.8 teeth, median 1.0) (p = 0.02), and higher attachment loss (mean 4.3 mm vs. 3.4 mm; median 4.0 mm vs. 3.0 mm) (p = 0.01). No major differences could be detected between the two groups in the pattern of affected teeth in the dentition.. The study demonstrates increased periodontal destruction among EOP-patients culture-positive for A. actinomycetemcomitans of the JP2-type compared with EOP-patients without the JP2-clone.

Topics: Actinobacillus Infections; Adolescent; Adult; Age Factors; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Bacterial Toxins; Base Pairing; Clone Cells; Cross-Sectional Studies; Dental Plaque; Exotoxins; Gene Deletion; Humans; Morocco; Periodontal Attachment Loss; Polymerase Chain Reaction; Statistics, Nonparametric

The aim of the present study was to investigate the intraoral distribution of Actinobacillus actinomycetemcomitans in young adults with minor signs of periodontal disease but harboring the organisms in the oral cavity. 17 healthy volunteers, 20 to 27 years of age, participated. Samples from mucosal surfaces of the oro-pharyngeal cavity and saliva (n = 221) as well as subgingival plaque from every tooth (n =477) were selectively cultivated for A. actinomycetemcomitans. Species identity and presence of the leukotoxin encoding gene, ltxA, were checked by multiplex polymerase chain reaction. Moreover, the leukotoxin promoter region was analyzed. No isolate harbored a 530 bp deletion in the promoter region of the leukotoxin gene, signaling minimally toxic strains. 42.1 +/- 30.4% extracrevicular and 34.4 +/- 29.5% subgingival samples were culture-positive. In extracrevicular samples, the organism could easily be recovered from cheek mucosa (62%), saliva (59%) and the palatal tonsils (41%). Mean log-transformed numbers of A. actinomycetecomitans colony forming units (CFU/ml) in culture-positive material ranged between 1.8 from the hard palate and 2.3 from 10 microl saliva. The highest prevalence in subgingival plaque was observed at maxillary 3rd molars (55%) followed by maxillary lateral incisors (50%) and mandibular 3rd molars (41%). Mean log-transformed counts of CFU/ml ranged between 2.2 at maxillary 3rd molars and 3.4 at upper central incisors. When adjusted for jaw, site and tooth type, the odds of isolating higher numbers of the organism were increased with every mm probing depth by a factor of 1.35 (p <0.05). The odds ratio for bleeding on probing was 1.38. Thus, in this young adult population with minor periodontal disease, A. actinomyetemcomitans was mainly associated with some deviation from gingival health. Of concern might be a minority of subjects (29%) with an extremely wide distribution of the organism in the oral cavity.

Topics: Adolescent; Adult; Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Base Pairing; Colony Count, Microbial; Dental Plaque; Exotoxins; Female; Gingiva; Gingival Hemorrhage; Humans; Incisor; Male; Molar, Third; Mouth; Mouth Mucosa; Odds Ratio; Oropharynx; Palatine Tonsil; Periodontal Diseases; Polymerase Chain Reaction; Prevalence; Promoter Regions, Genetic; Saliva; Sequence Deletion; Statistics as Topic; Tongue

The aim of this study was to determine the prevalence and the structure of the leukotoxin promoter region of Actinobacillus actinomycetemcomitans in an ethnic Chinese population.. Subgingival plaque samples were collected from 42 patients with moderate to advanced periodontitis and 50 periodontally healthy patients. A. actinomycetemcomitans was detected directly from the crude subgingival plaque by PCR using leukotoxin gene specific primers. The presence of A. actinomycetemcomitans was determined by a single 285 bp PCR amplicon.. A. actinomycetemcomitans was found to be present in the subgingival plaque of 68 out of a total of 92 patients examined (74%). 29 out of the 42 periodontitis patients tested were carriers of A. actinomycetemcomitans (69%). Among the periodontally healthy patients studied, 39 out of 50 subjects possessed the bacteria (78%). PCR analysis of the promoter region of the ltx operon revealed that none of the 42 moderate to advanced periodontitis patients examined harboured A. actinomycetemcomitans strains with the JP2-like promoter of the ltx operon, known to enhance leukotoxin expression. 2 out of the 27 advanced periodontitis patients clinically diagnosed as suffering from rapidly progressive periodontitis were found to be carriers of the mildly toxic strain of A. actinomycetemcomitans with the characteristic 652-like promoter.. The high prevalence of A. actinomycetemcomitans, regardless of whether the subgingival samples were analysed from patients with healthy or diseased periodontium suggests that this bacterial species is part of the normal oral flora of ethnic Chinese. Our preliminary results also suggested that subjects who harboured the mildly toxic strain of A. actinomycetemcomitans were potentially susceptible to aggressive forms of periodontitis.

Topics: Actinobacillus Infections; Adult; Aggregatibacter actinomycetemcomitans; Case-Control Studies; China; Dental Plaque; Exotoxins; Female; Gene Expression Regulation, Bacterial; Humans; Male; Middle Aged; Periodontitis; Polymerase Chain Reaction; Prevalence; Singapore

Actinobacillus actinomycetemcomitans leukotoxin is thought to be an important virulence factor in the pathogenesis of localized juvenile and other forms of early-onset periodontitis. Some highly leukotoxic A. actinomycetemcomitans strains produce 10 to 20 times more leukotoxin than other minimally leukotoxic strains. The distribution, clonality, and intrafamilial transmission of highly leukotoxic A. actinomycetemcomitans were examined in order to determine the importance of leukotoxin in the pathogenesis of periodontitis.. The polymerase chain reaction (PCR) was used to differentiate highly leukotoxic from minimally leukotoxic strains in examining 1,023 fresh A. actinomycetemcomitans isolates and strains from our culture collection. These were obtained from 146 subjects including 71 with localized juvenile periodontitis (LJP), 4 with early-onset periodontitis, 11 with post-localized juvenile periodontitis, 41 with adult periodontitis, and 19 periodontally normal subjects. The arbitrarily primed polymerase chain reaction (AP-PCR) analysis of 30 oral isolates from each of 25 subjects was used to determine the intraoral distribution of A. actinomycetemcomitans clones. AP-PCR was also used to examine the transmission of A. actinomycetemcomitans in 30 members of 6 families. The clonality of 41 highly leukotoxic A. actinomycetemcomitans strains was evaluated by both AP-PCR and ribotyping.. Highly leukotoxic A. actinomycetemcomitans was found only in subjects with localized juvenile and early-onset periodontitis. Fifty-five percent of the LJP subjects harbored highly leukotoxic A. actinomycetemcomitans isolates. Seventy-three percent of the A. actinomycetemcomitans isolates in these subjects were highly leukotoxic. Highly leukotoxic A. actinomycetemcomitans infected younger subjects (mean age 13.95 years, range 5 to 28 years) than minimally leukotoxic (mean age 35.47 years, range 6 to 65 years). Most subjects were infected with only one A. actinomycetemcomitans genotype. However, PCR of whole dental plaques and subsequent analysis of up to 130 individual oral isolates suggested a possible shift in A. actinomycetemcomitans over time in that a few subjects harbored both highly leukotoxic and minimally leukotoxic strains. AP-PCR analysis was consistent with intrafamilial A. actinomycetemcomitans transmission. Ribotyping and AP-PCR analysis confirmed a previous report that highly leukotoxic A. actinomycetemcomitans consists of a single clonal type.. This study suggests that localized juvenile and other forms of Actinobacillus-associated periodontitis are primarily associated with the highly leukotoxic clone of A. actinomycetemcomitans.

Topics: Actinobacillus Infections; Adolescent; Adult; Age Factors; Aged; Aggregatibacter actinomycetemcomitans; Aggressive Periodontitis; Bacterial Toxins; Chi-Square Distribution; Child; Child, Preschool; Clone Cells; Cytotoxins; Dental Plaque; Exotoxins; Female; Humans; Male; Middle Aged; Periodontitis; Periodontium

Actinobacillus actinomycetemcomitans produces a pore-forming leukotoxin that lyses human polymorphonuclear leukocytes and monocytes. Certain proteolytic bacteria may coexist with A. actinomycetemcomitans in periodontal pockets. We aimed therefore to examine whether oral bacteria can modify the leukotoxicity of A. actinomycetemcomitans. A total of 55 strains representing 45 bacterial species of the subgingival flora were tested. Each strain was incubated with the highly toxic strain of A. actinomycetemcomitans HK 1519 and the leukotoxic activity of the suspension against human polymorphonuclear leukocytes was determined from the activity of the lactate dehydrogenase released upon lysis of the leukocytes. Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica and Prevotella loeschii inhibited the leukotoxicity of A. actinomycetemcomitans cells as well as the activity of leukotoxin purified from the same strain. The bacterial strains without the ability to block leukotoxic activity also failed to destroy pure leukotoxin even after 5 h of incubation. The proteolytic degradation of leukotoxin by P. gingivalis was mainly dependent on the activity of the enzymes R- and K-gingipains. P. intermedia and P. nigrescens also degraded the leukotoxin by enzymes. The results imply a role of the periodontal microflora in modifying the virulence of A. actinomycetemcomitans by destroying its leukotoxin.

Topics: Adhesins, Bacterial; Aggregatibacter actinomycetemcomitans; Culture Media, Conditioned; Cysteine Endopeptidases; Dental Plaque; Ecosystem; Electrophoresis, Polyacrylamide Gel; Exotoxins; Gingipain Cysteine Endopeptidases; Hemagglutinins; Humans; Neutrophils; Porphyromonas gingivalis; Prevotella; Virulence

In a previous population-based study of 3896 7-9-year-old children living in Sweden, it was found that 32 children (0.8%) exhibited radiographic, periodontal bone loss at > or = 2 proximal surfaces of their deciduous teeth. In the present study, 26 of the 32 children were subjected to additional oral and systemic health examination. 20 other children without any radiographic evidence of bone loss in their primary dentition served as referents. None of the cases or the referents were detected to have any systemic disease. The frequency of bleeding and suppuration on probing, radiographic proximal calculus and probing attachment loss was higher among the cases than the referents. Actinobacillus actinomycetemcomitants was found subgingivally in 14 of the cases but in none of the referents. 11 of 22 cases analysed for presence of serum antibodies against A. actinomycetemcomitans leukotoxin were sero-positive compared to none of 7 referents available for analysis. Evaluation of the data from each child revealed wide variations in clinical parameters among the children in the case group. In this group, there were children with deep probing depths, probing attachment loss, suppuration on probing, proximal calculus and presence of subgingival A. actinomycetemcomitans, indicating current periodontitis. However, in the case group there were also children without positive signs of inflammatory disease, similar to the children in the reference group. In fact, the findings suggest that less than half of the number of individuals with > or = 2 proximal sites with bone loss had current periodontitis.

Topics: Aggregatibacter actinomycetemcomitans; Alveolar Bone Loss; Antibodies, Bacterial; Bacterial Toxins; Case-Control Studies; Child; Dental Calculus; Dental Plaque; Exotoxins; Female; Gingival Hemorrhage; Health Status; Humans; Male; Periodontal Attachment Loss; Periodontal Pocket; Periodontitis; Suppuration; Tooth, Deciduous

The purpose of this study was to assess the sensitivity and specificity of the PCR in detecting Actinobacillus actinomycetemcomitans. The PCR's detection capability was compared with those of three other methods: culture-enhanced PCR (CE-PCR), colony hybridization (CH), and conventional culture with presumptive biochemical identification. A 285-bp stretch of the leukotoxin gene lktA of A. actinomycetemcomitans was amplified by PCR with primers TT-15 and TT-16. For CH, the PCR product was labeled with digoxigenin and used as a hybridization probe. Nucleotide sequence analysis of the PCR product of A. actinomycetemcomitans 1D4 and 1664 and three clinical isolates revealed complete homology among the tested strains, with only one base substitution (at position 1344) in comparison with the published sequence. With artificially infected subgingival plaque, the detection limit of PCR for A. actinomycetemcomitans was 10(3) CFU/ml of plaque suspension. Culturing subgingival plaque on tryptic soy-serum-bacitracin-vancomycin agar prior to PCR (CE-PCR) improved the limit of detection to 10(2) CFU/ml. Analysis of subgingival plaque samples from 35 patients with periodontal disease and 10 periodontally healthy subjects revealed that CE-PCR and CH had the highest overall rate of A. actinomycetemcomitans detection (both 58%), followed by PCR and culture (both 42%). With CH as the "gold standard", the sensitivities of CE-PCR, PCR, and culture were 88, 65, and 58%, respectively; the specificities were 84, 89, and 79%, respectively. The CE-PCR provided acceptable positive and negative predictive values (> or = 70%) when the prevalence of A. actinomycetemcomitans varied between 30 and 70%. PCR alone provided comparable predictive values over a narrower range of prevalence rates (30 to 50%), while culture did not afford acceptable predictive values at any prevalence rate. PCR and CE-PCR were found to be superior to culture with presumptive biochemical identification and should be the preferred methods for the detection of A. actinomycetemcomitans in subgingival plaque.

Topics: Aggregatibacter actinomycetemcomitans; Bacterial Toxins; Bacteriological Techniques; Base Sequence; Dental Plaque; DNA Primers; DNA, Bacterial; Exotoxins; Gingiva; Humans; Molecular Sequence Data; Polymerase Chain Reaction; Sensitivity and Specificity

The effects of heat labile, high molecular weight water-soluble toxins from bacterial plaque on HL60 promyelocytic cells were examined. On gel filtration, four inhibitors of HL60 cell growth and two inhibitors of HeLa cell growth (PT1, PT2) were detected. The first and third HL60 cell inhibitors corresponded to the two HeLa cell inhibitors. The last eluted HL60 cell inhibitor (plaque leukotoxin, PL) did not inhibit HeLa cell growth. Anti-PT2 antibodies reduced the activity of enriched PT2 by 20-50%, but all other antisera tested exhibited no effect. Anti-PL antibodies detected antigens from Actinobacillus actinomycetemcomitans, although anti-A. actinomycetemcomitans and anti-Capnocytophaga sputigena antibodies did not react with plaque extract. These findings suggest that the plaque toxins examined in this study were probably not derived from these two bacteria.

Topics: Aggregatibacter actinomycetemcomitans; Animals; Antibodies, Bacterial; Bacterial Toxins; Capnocytophaga; Chromatography, Gel; Chromatography, Ion Exchange; Cross Reactions; Dental Plaque; Exotoxins; HeLa Cells; HL-60 Cells; Humans; Immunosuppressive Agents; Rabbits

Topics: Actinobacillus; Adolescent; Adult; Aggressive Periodontitis; Animals; Cell Survival; Dental Plaque; Exotoxins; Humans; Macaca fascicularis; Male; Microscopy, Electron; Mitogens; Monocytes; Neutrophils

Topics: Actinobacillus; Adhesiveness; Bacteroides; Capnocytophaga; Chemotaxis, Leukocyte; Cytophagaceae; Dental Plaque; Ecology; Exotoxins; Humans; Immunoglobulins; Periodontal Diseases; Phagocytosis; Virulence