Cryptosporidiosis is a very important opportunistic infection and is responsible for significant morbidity and mortality in HIV/AIDS patients. Although current laboratory methods are generally considered adequate to detect high concentrations of oocysts, they fail to detect cases of cryptosporidiosis in many immunocompromised patients. The present study was done to determine the diagnostic efficacy of modified Ziehl-Neelsen (ZN), antigen detection ELISA, and a nested PCR assay for detection of Cryptosporidium in 58 adult AIDS cases with diarrhea from the ART clinic of Lok Nayak Hospital, New Delhi. Cryptosporidium was detected in 17 (29.4%), 39 (67.3%), and 45 (77.5%) cases by modified ZN staining, antigen ELISA, and nested PCR assay, respectively. Taking nested PCR as the gold standard, specificity of both modified ZN staining and Cryptosporidium antigen detection ELISA was 100% while the sensitivity of the tests was 37.8% and 86.6%, respectively. PCR was more sensitive than the other two diagnostic modalities but required a more hands-on time per sample and was more expensive than microscopy. PCR, however, was very adaptable to batch analysis, reducing the costs considerably. This assay can therefore have considerable advantages in the treatment of immunosuppressed individuals like AIDS patients, allowing their early diagnosis and decreasing the morbidity and the mortality. 1. Introduction Cryptosporidium parvum is an enteric protozoan parasite with worldwide distribution which may inhabit the gastrointestinal tract of a wide variety of animals including humans [1, 2]. Infection with C. parvum in immunocompetent persons often results in asymptomatic or mild self-limited disease, but in HIV-infected patients, particularly those with low CD4 counts, infection may result in chronic or life-threatening diarrhea, or extraintestinal disease [3–6]. In developed countries, an estimated 14% of AIDS patients with diarrhea have C. parvum infection [3]. In developing countries, the parasite is reported in 24% (range: 8.7–48%) of HIV-seropositive adults and children while in India it has been reported in 2–60% of such patients [7–11]. In persons with AIDS, cryptosporidiosis is commonly a permanent diarrheal illness that leads to chronic malabsorption of fluids, nutrients, vitamins, and electrolytes with resulting wasting [12]. This warrants a prompt diagnosis and an early institution of specific therapy to reduce the morbidity associated with the disease. But over the years, detection of this protozoon has become a challenge. Cryptosporidiosis is most commonly
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