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AWWA WQTC52765
- Detect Infectious Cryptosporidium parvum Oocysts Using Integrated Cell Culture-rRNA In Situ Hybridization
- Conference Proceeding by American Water Works Association, 01/01/2000
- Publisher: AWWA
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In an infectivity assay, human illeocecal adenocarcinoma (HCT-8) cellswere infected with different concentrations of Cryptosporidium parvum oocysts. After 48 hours of incubation, in-situ hybridization (ISH) using digoxigenin-labeled DNA probes targeted at the C. parvum-specific small subunit rRNA sequences with colorimetric detection was performed to detect infectious oocysts. In addition, immunofluorescencedetection of intracellular reproductive stages of C. parvum using fluorescein-labeled polyclonal antibody against sporozoites of C. parvum was conducted and compared with ISH. Both C. parvumand C. muris were used to infect HCT-8 cells along with ISH to determine the specificity of the DNA probes. Includes 18 references, tables, figures.