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746 GENETIC VARIATION IN CRYPTOSPORIDIUM PARVUM ISOLATES FROM CHILDREN WITH DIARRHEA IN BANGLADESH. Kirthi Hira, Wasif A. Khan, Melanie R. Mackay, Seth Cohen, Brett A. Leav, Stephen B. Calderwood, Edward T. Ryan, Honorine D. Ward. Division of Geographic Medicine and Inectious Diseases, Tufts-New England Medical Center, Boston, MA; International Centre for Diarrheal Diseases Research, Dhaka, Bangladesh; Division of Infectious Diseases, Massachussetts General Hospital, Boston, MA; Africa Centre for Health and Population Studies, Mtubatuba, South Africa. Very little is known about the molecular epidemiology of cryptosporidiosis, particularly in developing countries. Genetic analysis has identified two major genotypes of Cryptosporidium parvum. Cp ; , the species which most commonly infects humans. Genotype I isolates infect humans exclusively, while genotype II isolates infect both humans and animals. Recent studies have identified significant intragenotypic heterogeneity among isolates. Extensive polymorphisms have been identified at the Cpgp40 15 locus, particularly among genotype I isolates. Based on these polymorphisms, thus far five allelic subgroups have been identified among genotype I isolates and one among genotype II isolates The goal of this study was to determine the species and genotype of isolates from 46 Cp-infected children in Bangladesh, to analyze Cpgp40 15 polymorphisms and to correlate genetic information with epidemiological and clinical parameters. DNA was extracted from 45 of the 46 stool samples and used for genetic analysis. PCR-RFLP analysis at the small-subunit rRNA locus was used for species determination. Genotyping was performed by PCR-RFLP analysis of the thrombospondin-related adhesive protein of Cp and Cp oocyst wall protein. Cpgp40 15 sequences were PCR-amplified from Cp DNA using primers derived from conserved regions. All 45 isolates were confirmed to be Cp. 40 of the isolates 89% ; were genotype I. No significant differences were detected between genotype and clinical or epidemiological parameters. Cpgp40 15 sequences could be PCR amplified from 41 of the isolates and the PCR products are being sequenced and classified into allelic subgroups. These results are consistent with those of previous studies from developing countries, which have shown Cp to be the species most commonly identified, and genotype I to be the most predominant genotype. Analysis of Cpgp40 15 polymorphisms, classification into allelic subgroups and correlation of genetic with clinical and epidemiological parameters in larger numbers of isolates may contribute to our understanding of the molecular epidemiology of cryptosporidiosis in this population.

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