ABSTRACT
Despite the availability of many parasitological methods for detection of Cryptosporidium and Isospora (Cystoisospora) belli in fecal samples, there are uncertainties about the accuracy of these techniques in laboratory practice. In this study, 27 formalin-fixed positive stool samples for Cryptosporidium and 15 for I. belli were analyzed by 2 concentration methods, sedimentation by centrifugation (SC) and formalin-ethyl acetate (FE), and by 3 tintorial techniques, modified Ziehl-Neelsen (ZN), safranin (SF), and auramine (AR). No significant differences were observed on Cryptosporidium identification between concentration methods, while a significantly higher number of I. belli oocysts (P < 0.0001) was detected in fecal smears concentrated by the SC than by the FE method. Fecal samples processed by FE produced a median oocyst loss to the fatty ring of 34.8% for Cryptosporidium and 45.4% for I. belli. However, FE concentration provided 63% of Cryptosporidium and 100% of I. belli slides classified as superior for microscopic examination. Regarding the efficiency of staining methods, a more significant detection of Cryptosporidium oocysts was observed in fecal smears stained by ZN (P < 0.01) or AR (P < 0.05) than by the SF method. Regular to high-quality slides for microscopic examination were mostly observed in fecal smears stained with AR or ZN for Cryptosporidium and with SF or ZN for I. belli. This study suggests a great variability in oocyst power detection by routine parasitological methods, and that the most frequent intestinal coccidians in humans have specific requirements for concentration and staining.
Subject(s)
Cryptosporidiosis/diagnosis , Cryptosporidium/isolation & purification , Feces/parasitology , Isospora/isolation & purification , Isosporiasis/diagnosis , Acetates , Acquired Immunodeficiency Syndrome/complications , Benzophenoneidum , Centrifugation/methods , Coloring Agents , Cryptosporidiosis/parasitology , Diarrhea/parasitology , Fixatives , Formaldehyde , Humans , Indicators and Reagents , Isosporiasis/parasitology , Phenazines , Staining and Labeling/methodsABSTRACT
To compare the efficacy of stool examination for the detection of Strongyloides stercoralis and hookworm, a total of 634 stool samples from the routine laboratory service of the Pharmacia Faculty, Federal University of Bahia, Brazil, were examined by agar plate culture (APC), Baermann-Moraes and spontaneous sedimentation. The sensitivity of agar plate culture, calculated by combining results of all 3 methods, was 95% for S. stercoralis and 77.6% for hookwoorm. Moreover, APC had superior accuracy than Baermann-Moraes and spontaneous sedimentation for S. stercoralis and hookworm diagnosis, respectively. The S. stercoralis and hookworm positive samples from the laboratory routine, obtained after the previous analysis, along with those initially selected, were used to evaluate the concordance between microscopic examination and both the type of furrows left by larvae and the time for culture positivity using the APC method. Of 115 stool samples positive for S. stercoralis and 92 positive for hookworm, 110 (95.7%) and 89 (96.7%), respectively, had concordant results for furrows and morphological characteristics. The cumulative percentage of positivity increased to 94% by the third day of observation; at this time, only 19.6% of hookworm-positive samples had positive culture plates. Analyses of 74 S. stercoralis-positive stool samples stored at 4°C for 24, 48 and 72h showed the presence of larvae in 48.6%, 28.4% and 23% of samples, respectively when re-examined by the APC. As a definitive diagnosis of strongyloidiasis depends on the microscopic demonstration of parasites, increasing the sensitivity of the detection requires the use of different parasitological methods, including APC.
