4-4--bis(2-di(2-hydroxyethyl)amino-4-(3-sulphophenylamino)-1-3-5-triazine-6-ylamino)stilbene-2-2--disulphonic-acid and Intestinal-Diseases--Parasitic

Intestinal microsporidiosis is the most common cause of chronic diarrhea in HIV/AIDS infected patients. The diagnosis of intestinal microsporidia depends on the detection of the spores by staining either with Chromotrope 2R or with fluorchrome uvitex 2B methods.. To compare the Chromotrope-2R and Uvitex-2B in detecting intestinal microsporidial spores from HIV/ AIDS patients at Nekempte Hospital.. A total of 120 single fresh stool samples were collected, and processed by water ether sedimentation method; stained with Uvitex-2B and observed microspridial spore under fluorescent microscope. From same stool samples, smear were prepared and stained with Chromotrope-2R method for the detection of intestinal microsporidial spores using light microscope.. Uvitex 2B detected 5/120 (4.2%) while Chromotrope 2R detected 4/120 (3.3%) and there was no statistical significance difference between the two methods (P>0.05). The sensitivity and specificity of the chromotrope-2R method relative to Uvitex-2B were 80% and 100%, respectively and positive and negative predictive values of Chromotrope-2R relative to the Uvitex 2B were 100% and 99%, respectively.. Based on its relative simplicity for processing, in terms of low cost materials (light microscopes compared to fluorescent microscopes) and reagents, make Chromotrope-2R to be recommended for diagnosis of microsporiadia infection in peripheral labs. Even though Uvitex-2B is superior, its application in peripheral health facilities is questionable and demanding.

Topics: Adolescent; Adult; AIDS-Related Opportunistic Infections; Benzenesulfonates; Coloring Agents; Cross-Sectional Studies; Ethiopia; Female; Humans; Intestinal Diseases, Parasitic; Male; Microsporidiosis; Naphthalenesulfonates; Sensitivity and Specificity; Spores, Fungal; Staining and Labeling

At present, the laboratory diagnosis of intestinal infections caused by microsporidia depends on the detection of the typical spores either with a modified trichrome stain (MTS) or by staining with fluorochromes. The purpose of the present study was (i) to compare staining with MTS (MTS method) and the staining with the fluorochrome Uvitex 2B (U2B method) with respect to their sensitivities and specificities, particularly in the presence of low numbers of spores, and (ii) to evaluate their reliabilities under routine laboratory conditions. First, 30 negative human stool specimens as well as 30 specimens enriched with a low concentration of microsporidial spores were examined. The U2B and MTS methods detected 27 and 30, of the positive samples, respectively (95% confidence intervals for sensitivity, 0.73 to 0.98 for the U2B method and 0.88 to 1.00 for the MTS method) without yielding false-positive results (95% confidence intervals for specificity, 0.88 to 1.00 for the MTS and U2B methods). In addition, analysis of serial dilutions of 17 stool specimens from AIDS patients containing microsporidia revealed comparable detection thresholds (P = 0.52) for both methods. Finally, 40 slides prepared from one stool specimen containing very few microsporidia and 40 negative slides were included in the routine diagnostic program during 1 month in order to monitor laboratory handling and run-to-run variations. Again, both methods exhibited comparable sensitivities (95% confidence intervals, 0.83 to 0.99 for the MTS method and 0.91 to 1.00 for the U2B method) and specificities (95% confidence intervals, 0.91 to 1.00 for the MTS and U2B methods). In conclusion, MTS and U2B methods are equally useful in the diagnosis of microsporidiosis. However, since detection thresholds for both methods differed considerably in all diluted stool specimens, performance of a combination of both methods may be more sensitive than the performance of only one procedure in the event of very low numbers of microsporidial spores.

Topics: AIDS-Related Opportunistic Infections; Animals; Azo Compounds; Benzenesulfonates; Eosine Yellowish-(YS); Feces; Humans; Intestinal Diseases, Parasitic; Methyl Green; Microsporida; Microsporidiosis; Sensitivity and Specificity; Specimen Handling; Spores; Staining and Labeling

Fluorescent stains with Uvitex 2B or other fluorochromes are widely used today, especially for the diagnosis of microsporidian infections in human immunodeficiency virus (HIV)-infected patients. Our objective was to ascertain whether the fluorescent stain with Uvitex 2B is also able to detect Isospora belli in stool samples and duodenal/bile juice of HIV-infected patients.. Case study.. University hospital of the University of Cologne, Germany.. Two HIV-infected patients with chronic diarrhea who had intestinal infections with I. belli.. Stool was concentrated by a modified water-ether sedimentation method, and duodenal/bile juice was concentrated by centrifugation. Wet-mount preparations were examined by phase-contrast and bright-field microscopy, and smears were stained with a modified acid-fast stain and a fluorescent stain with Uvitex 2B.. Using the fluorescent stain with Uvitex 2B, the oocysts of I. belli stained bright white/blue fluorescent and showed a structure similar to that of the oocysts in acidfast stains.. Staining with Uvitex 2B is a suitable method for the diagnosis of I. belli infections. This technique can be used for the diagnosis of three important gastrointestinal parasites (viz, microsporidia, cryptosporidia, and I. belli) responsible for diarrhea in HIV-infected patients.

Topics: Adult; AIDS-Related Opportunistic Infections; Animals; Benzenesulfonates; Bile; Feces; Female; Fluorescent Dyes; Humans; Intestinal Diseases, Parasitic; Isospora; Isosporiasis; Male; Staining and Labeling