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AWWA QTC97116
- Detection of Cryptosporidium parvum by Enzyme Immunoassay and the Polymerase Chain Reaction
- Conference Proceeding by American Water Works Association, 01/01/1997
- Publisher: AWWA
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The detection of Cryptosporidium parvum oocysts in environmental water samples and the determination of oocyst infectivity remain pressing issues for the water industry. In this study, the polymerase chain reaction (PCR) with primers specific for the C. parvum hsp70 gene were used to detect total C. parvum oocysts and a cell culture (HCT-8) enzyme-linked immunosorbent assay (ELISA) was used to detect infectious oocysts. An integrated cell culture-PCR assay was also evaluated for its use as a highly sensitive C. parvum infectivity assay. Oocysts seeded into environmental water samples were recovered by immunomagnetic separation (IMS) and used directly for PCR analyses and inoculation of HCT-8 cell cultures. PCR on oocyst stocks revealed a consistent detection limit of 1 to 2 oocysts. ELISA alone had a detection limit between 10 and 100 oocysts, while the integrated cell culture-PCR had a detection limit of 5 or less infectious oocysts in a background of 1,000,000 HCT-8 cells.
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