• AWWA WQTC50425
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AWWA WQTC50425

  • Finished Water Monitoring for Infectious Cryptosporidium Oocysts Using Immunomagnetic Separation (IMS) and Cell Culture-Polymerase Chain Reaction Method (CC-PCR)
  • Conference Proceeding by American Water Works Association, 01/01/1999
  • Publisher: AWWA

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A highly specific and sensitive diagnostic technique to detect live infectious Cryptosporidium parvum oocysts which is based on the use of cell cultures and DNA amplification by PCR (CC-PCR), was applied to monitor finished water for contamination with live infectious oocysts. The experimental design considered the analysis of blind samples from 25 treatment plants located in eight states. 102 finished water samples were collected with a final volume of 100¿5 L, using the Envirochek Sampling Capsule. The sample processing at the Belleville Laboratory initiated with sample elution which was carried out as described by the US Environmental Protection Agency 1622 method with slight modifications. After the immunomagnetic separation of oocysts (IMS), all purified concentrates were submitted to CC-PCR assay. Within the monitoring, less than 1.0% of the samples tested positive for live infectious C. parvum. DNA sequence analysis of the hsp70 gene proved that the genotype from the in vitro-cultured specimen was 100% identical to the bovine C. parvum genotype. Due to the high frequency of waterborne human cryptosporidiosis outbreaks, this study provides evidence of the survival and infectivity of stressed oocysts after chemical treatment in finished drinking water. Therefore, the practical applications of the methodology described in this study could help ensure the quality of finished drinking water and ultimately benefit the water industry. Includes 6 references, tables.

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