Research in Medicine

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Background: Cryptosporidiosis is prevalent in animals and human beings, both immuno-competent and immuno-compromised persons, worldwide. Epidemiologically, it is important to differentiate between human and animal species. The aim of this study is to define the utility of a 1055 bp fragment of 18s rRNA for differentiation of Human and Cattle Cryptosporidiosis. Material and methods: 1020 human fecal samples and 940 bovine fecal samples were collected and tested for Cryptosporidium by modified acid fast staining. DNA of positive samples was extracted and a 1055bp fragment of 18s rRNA was amplified by Nested-PCR technique using 4 designed primers. The second PCR product was digested by Ssp1&Vsp1 restriction enzymes to determine the Cryptosporidium species and the results were analyzed by 2.5% agarose gel electrophoresis and visualized after ethidium bromide staining. Results: 12 human samples and 23 cattle samples were detected as being positive for Cryptospridium by modified acid fast method. These results were confirmed by molecular technique. All cattle samples and 10 human samples presented similar bands on 2.5% agarose gel which confirmed Cryptosporidium parvum except two human samples which had different patterns. On sequencing these 2 samples these two new isolates were found to be different from the rest. The sequences of 2 new human isolates were different from others in Gene Bank so they were deposited in Gene Bank at accession No: EU311203 Conclusion: In this study we detected both cryptosporidium parvum & cryptosporidium hominis in human samples but cryptosporidium parvum was more prevalent.

Keywords: Cryptosporidium, RNA, Polymerase Chain Reaction.

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