Improvement and application of Cryptosporidium detection with Florescent in situ Hybridization

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Fluorescence in situ hybridization (FISH) was applied for specific detection and distinction of Cryptosporidium oocysts by microscopic observation. High fluorescence was observed using the DIAG-CRYR1 probe targeting 18S rRNA partial sequences to distinguish Cryptosporidium spp from Cryptosporidium parvum HNJ-1 oocysts. Combining FISH with fluorescence antibody staining, DAPI, and differential interference contrast observation was valuable for easy differentiation of Cryptosporidium oocysts by microscopic observation. Determination of Cryptosporidium variability by FISH was conducted. Heat inactivated oocysts declined FISH stainability until 100 hours in both PBS and filtrated sewage.