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A MULTIPLEX REVERSE TRANSCIPTION-PCR METHOD FOR DETECTION OF HUMAN ENTERIC VIRUSES IN GROUNDWATER
Citation:
Fout, G S., B C. Martinson, M. N. Moyer, D R. Dahling, AND L J. Wymer. A MULTIPLEX REVERSE TRANSCIPTION-PCR METHOD FOR DETECTION OF HUMAN ENTERIC VIRUSES IN GROUNDWATER. APPLIED AND ENVIRONMENTAL MICROBIOLOGY 69(6):3158-3164, (2003).
Impact/Purpose:
Overarching Objectives and Links to Multi-Year Planning
This task directly supports the Drinking Water Research Program Multi-Year Plan's long term goal to "develop scientifically sound data and approaches to characterize and manage risks to human health posed by exposure to waterborne pathogens and chemicals" under GRPA Goal 2 (Clean and Safe Water). The overarching objective is to provide the Office of Water, Agency risk assessors and managers, academics, the scientific community, state regulators, water industry and industry spokes groups with exploratory occurrence and exposure data on human enteric viruses. These data will improve the quality of risk-based assessments and tools used by the Agency to set regulations, policies and priorities for protecting human health and allow the Agency to assure the public that the appropriate methods are being used to demonstrate that drinking water is safe from pathogenic agents.
Specific Subtask Objectives:
o Conduct an exploratory occurrence studies on emerging human waterborne pathogenic viruses and viruses on the Contaminant Candidate List (CCL) in water (Subtask A; to be completed by 9/05 in support of LTG 1 (due 2010)).
o Determine the relationship of bacterial virus indicators to human enteric virus occurrence in the above studies (Subtask A; to be completed by 9/05 in support of LTG 1 (due 2010)).
o Develop a non-invasive assay for measuring human exposure to viruses (Subtask B; to be completed by 9/05 in support of LTG 1 (due 2010)).
Description:
Untreated groundwater is responsible for about half of the waterborne disease outbreaks in the United States. Human enteric viruses are thought to be leading etiological agents of many of these outbreaks, but there is relatively little information on the types and levels of viruses found in groundwater. To address this problem, monthly samples from 29 groundwater sites were analyzed for one year for enteroviruses, hepatitis A virus (HAV), Norwalk virus, reovirouses and rotaviruses by multiplex reverse transcription-PCR (RT-PCR). A procedure was developed to remove environmental inhibitors of RT-PCR from groundwater samples. The procedure allowed an average of 71 liters of the original groundwater to be asssayed per RT-PCR reaction with an average virus recovery rate of 74%, based upon seeded samples. Human enteric viruses were detected in 16% of the groundwater samples analyzed with reoviruses being the most frequently detected virus group.