Development of a Rapid, Quantitative Method for the Detection of Infective Coxsackie and Echo Viruses in Drinking WaterEPA Grant Number: R828040
Title: Development of a Rapid, Quantitative Method for the Detection of Infective Coxsackie and Echo Viruses in Drinking Water
Investigators: Yates, Marylynn V. , Chen, Wilfred , Mulchandani, Ashok
Institution: University of California - Riverside
EPA Project Officer: Nolt-Helms, Cynthia
Project Period: February 1, 2000 through February 1, 2002
Project Amount: $321,784
RFA: Drinking Water (1999) RFA Text | Recipients Lists
Research Category: Drinking Water , Water
The objectives of this research are to improve on the current analytical methods for quantitative detection of infective coxsackie and echo viruses in drinking water. The specific objectives of this research are to: (1) Improve the sensitivity and specificity of IMS-PCR for infective coxsackie and echo virus detection in water samples with the use of monoclonal antibodies; (2) Improve the efficiency and quantitation capabilities of IMS-PCR with the use of molecular beacons; and (3) Determine and quantify the presence and viability of coxsackie and echo viruses in concentrated drinking water samples.
We propose to combine IMS-PCR with molecular beacons to increase the sensitivity and specificity of analysis of environmental water samples for coxsackie and echo viruses. The research approach will include the: 1) optimization of IMS-PCR with monoclonal antibodies to the coxsackie and echo viruses; 2) development of fluorogenic probes for the target viruses; 3) analysis of virus-seeded water with the IMS - molecular beacon combination, and 4) testing of protocols on seeded drinking water concentrates to enable an assessment of the sensitivity of the method to inhibitors present in environmental waters. The specificity and quantitative capability of the protocols will be assessed by comparing results to quantitative cell culture analysis. All experiments will also be conducted with poliovirus to provide a basis for comparison.
The lack of standardized methods that can be routinely performed to detect and quantify infective coxsackie and echo viruses has limited the amount of information available on the occurrence of these viruses in drinking water and other environmental samples. Thus, an assessment of the risk of coxsackie and echo virus infection as a result of exposure to drinking water is not possible at this time. The methods developed in this study should improve our ability to provide quick and efficient results for the detection and quantitation of coxsackie and echo viruses in samples from environmental waters. This will enable the collection of adenovirus occurrence data in drinking water, which can then be used to assess the potential public health risks from these organisms. In addition, the methods can be adapted to facilitate the detection of other microorganisms in water, improving our ability to calculate risks from those organisms as well.