Blind trials were performed on an optimized cell culture-immunofluorescence (IFA)procedure, using the HCT-8 cell line, to determine the sensitivity and reproducibility formeasuring infectivity of flow cytometry prepared inocula of C. parvum oocysts. Usingbetween 0-100% viable oocysts, prepared at the U.S. Environmental Protection Agency (USEPA), cell culture-IFA wasperformed on known oocyst inocula of unknown infectivity at the American Water (AW) laboratory.Results indicated the control (100% live) oocyst suspensions yielded statistically similarresults to cell culture dose response curves developed previously at AW. For test samplescontaining oocyst suspensions of unknown infectivity, cell culture-IFA analyses indicateda high degree of correlation (r2= 0.89; n=26) with the values expected by USEPA. Cellculture infectivity correlates well with neonatal mouse infectivity assays and these "blind"validation trials provide credibility for the cell culture-IFA procedure as a cost-effectiveand expedient alternative to mouse infectivity assays for determining in vitro infectivityof C. parvum oocysts. Includes 20 references, figures.
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Edition: Vol. - No. Published: 11/01/2005 Number of Pages: 11File Size: 1 file , 290 KB