Genotyping of single Cryptosporidium oocysts isolated from sewage and river water
1Water Environment Laboratory, School of Environmental Health, Azabu University, 1-17-71 Fuchinobe, Sagamihara, 229-8501 Kanagawa, Japan. hiratat@azabu-u.ac.jp
Abstract:
The study was designed to genotype individual Cryptosporidium oocysts using an 18S rRNA gene-based semi-nested PCR and direct sequencing procedure. Positive PCR amplification was observed in all single C. parvum HNJ-1 oocyst samples tested. Semi-nested PCR and direct sequencing was applied to Cryptosporidium oocysts isolated from sewage and river water. The procedure could genotype 54% of FITC-stained single oocysts isolated from sewage and 32% from river water. The predominant genotype in both sewage and river water was C. parvum genotype 1, accounting for 33 and 25%, respectively, of all the FITC-stained intact Cryptosporidium oocysts present.
