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Applied and Environmental Microbiology, June 2003, p. 3158-3164, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3158-3164.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Multiplex Reverse Transcription-PCR Method for Detection of Human Enteric Viruses in Groundwater

G. Shay Fout,1* Beth C. Martinson,1 Michael W. N. Moyer,2 and Daniel R. Dahling1

National Exposure Research Laboratory, Office of Research and Development,1 Technical Support Center, Office of Water, U.S. Environmental Protection Agency, Cincinnati, Ohio 452682

Received 7 August 2002/ Accepted 19 March 2003

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 1 year for enteroviruses, hepatitis A virus, Norwalk virus, reoviruses, and rotaviruses by multiplex reverse transcription-PCR (RT-PCR). A procedure with which to remove environmental RT-PCR inhibitors from groundwater samples was developed. The procedure allowed an average of 71 liters of the original groundwater to be assayed per RT-PCR, with an average virus recovery rate of 74%, based on seeded samples. Human enteric viruses were detected in 16% of the groundwater samples analyzed, with reoviruses being the most frequently detected virus group.

* Corresponding author. Mailing address: U.S. EPA, NERL, 26 W. Martin Luther King Dr., Cincinnati, OH 45268-1320. Phone: (513) 569-7387. Fax: (513) 569-7117. E-mail: fout.shay{at}epa.gov.

Applied and Environmental Microbiology, June 2003, p. 3158-3164, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3158-3164.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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