Abstract

BackgroundEnteric protozoa and sporozoa have emerged as important opportunistic parasites and can cause fatal infections in AIDS patients. The line of treatment being different for them necessitates an accurate and prompt identification of these to avoid empirical treatment. In this study which is the first of its kind from India we did a comprehensive evaluation of different techniques, comparing them on the basis of the attributes like yield, cost, time taken, expertise and infrastructure. For the first time combination of Calcoflour White and DAPI, a nuclear stain, were used to identify Microsporidia spp. Thus, a diagnostic protocol was devised for rapid, sensitive and cost effective identification of the opportunistic enteric protozoa.ResultsThe organisms isolated from the stool samples of the cases (450 HIV patients) were predominantly Cryptosporidium spp., Microsporidia spp. and Cyclospora spp. Interestingly, the control group (200 relatives of the patients who were HIV negative) showed a high incidence (21%) of Cryptosporidium spp. We found a significant increase in the sensitivity of microscopy in detecting Cryptosporidium spp. and Cyclospora spp. after formol ether concentration. Kinyoun's staining was better compared to Modified safranin staining for Cryptosporidium spp. identification. Although ELISA had a sensitivity of 93.25% and specificity of 97% for Cryptosporidium spp. detection, we ranked Kinyoun's staining better than ELISA because it is not affordable to most of our patients. For detecting Cyclospora cayetanensis, autoflourescence was the easiest and most cost effective method followed by Safranin technique. Combination of Calcoflour White stain and DAPI gave good results for the identification of Microsporidia spp. We assessed the above techniques and graded the attributes in the following descending order: cost effectiveness, sensitivity, ease of use and interpretation, time taken for the procedure and batch testing.ConclusionThus, we conclude that a combination of minimum three procedures should be carried out for the screening of stool specimens of HIV positive patients. Kinyoun's staining should be made mandatory for every diarrheal stool sample from HIV patients. Also every laboratory should assign its own value to the attributes and apply Multiattribute utility theory or the Analytical hierarchy process to decide the most appropriate methodology.

Highlights

  • Enteric protozoa and sporozoa have emerged as important opportunistic parasites and can cause fatal infections in AIDS patients

  • We observed a high prevalence of Cryptosporidium spp. (21%) in the control group which comprised of HIV negative family members having diarrhea and coming from similar environmental, social and economic background as that of HIV patients

  • We found a significant increase in the sensitivity of microscopy in detecting Cryptosporidium spp. and Cyclospora spp. after formol ether concentration (Chi square test, p < 0.05)

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Summary

Introduction

Enteric protozoa and sporozoa have emerged as important opportunistic parasites and can cause fatal infections in AIDS patients. Coccidia and sporozoa like Cryptosporidium spp., Microsporidia spp., Isospora spp. and Cyclospora spp. have emerged as important parasites. Infection with these protozoa usually causes nausea, low grade fever, abdominal cramps, anorexia and watery motions [1]. Beginning from examination of small or large bowel biopsy material to different staining techniques and their modifications, several methods have been adopted. Many of these techniques are cumbersome and time consuming. Rapid and sensitive techniques are needed to give an early diagnosis of these protozoal infections as the results can influence therapeutic intervention

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