Development of a Quantitative Cell Culture-Based Infectivity Assay for Cryptosporidium parvum in Source and Finished Water

EPA Grant Number: R825146
Title: Development of a Quantitative Cell Culture-Based Infectivity Assay for Cryptosporidium parvum in Source and Finished Water
Investigators: Leon, Ricardo De , Rochelle, Paul A.
Institution: Metropolitan Water District of Southern California
EPA Project Officer: Hiscock, Michael
Project Period: October 1, 1996 through September 30, 1998
Project Amount: $214,502
RFA: Drinking Water (1996) RFA Text |  Recipients Lists
Research Category: Drinking Water , Water

Description:

Detection of Cryptosporidium oocysts in finished water is of concern because these protozoa have low infectious doses and high resistance to chlorination. Nonetheless, current detection methodology used for water samples, the immunofluorescent assay (IFA), does not determine whether the oocysts are viable or infectious. The purpose of this project is to develop a cell culture infectivity assay in combination with the rapid and specific methods of in-situ gene probe hybridization and polymerase chain reaction (PCR). The project objectives are: 1) optimize cell culture assay for infection with low levels of oocysts and develop a reverse transcriptase (RT)-PCR assay which targets Cryptosporidium parvum specific mRNA from a heat shock protein; 2) develop a quantitative in-situ (IS)-PCR cell culture-based infectivity assay for Cryptosporidium with confirmatory non-radioactive in-situ probing; 3) develop sample clean-up procedures which are compatible with cell culture and optimally isolate viable or infectious oocysts from environmental samples; and 4) evaluate the developed method with environmental concentrates from source and finished water. The expected outcome and benefits to public health and the water industry are the development of a quantitative method for determining the infectivity of C. parvum oocysts isolated from water. The assay will provide a better method to assess the risk to public health posed by C. parvum oocysts in finished water. It may also be used to determine the efficacy of oocyst inactivation by disinfectants and to monitor oocyst survival in water.

Publications and Presentations:

Publications have been submitted on this project: View all 7 publications for this project

Journal Articles:

Journal Articles have been submitted on this project: View all 7 journal articles for this project

Supplemental Keywords:

pathogens, drinking water, genetic probes, California, CA, region 9, exposure, environmental testing, human health., RFA, Scientific Discipline, Geographic Area, Water, Genetics, Environmental Chemistry, Health Risk Assessment, State, Analytical Chemistry, Drinking Water, cryptosporidium parvum oocysts, monitoring, microbial risk assessment, human health effects, exposure and effects, exposure, source water, infectivity assays, treatment, gene probe, emerging pathogens, drinking water contaminants, California (CA), pathogenic protozoa, immunofluorescent assay, drinking water system

Progress and Final Reports:

  • 1997
  • Final Report