Affiliation:
1. American Water Works Service Company, Inc., Belleville, Illinois 62220,1 and
2. American Water Works Service Company, Inc., Voorhees, New Jersey 080432
Abstract
ABSTRACT
We evaluated the use of the PCR for detection of enteric viruses in groundwater. To do this, we used an improved sample-processing technique and a large-volume amplification protocol. The objective of this study was to use advanced molecular techniques to develop a rapid and simple method which can be used by the water industry for detection of viral contamination in a variety of water samples. The strategy described here fulfills the water industry’s need for a rapid, reliable, easily performed method for analyzing groundwater for virus contamination. Viruses were detected after concentration from at least 400 gallons (1,512 liters) of water by a filter adsorption and elution method, which resulted in a concentrate containing viruses. A total of 150 samples were analyzed by performing cell culture assays for enteroviruses and by performing reverse transcription PCR (RT-PCR) analyses for enteroviruses, hepatitis A virus, and rotavirus. Thirteen samples (8.7%) produced cellular cytopathic effects when the Buffalo green monkey cell line was used. When primers specific for enteroviruses were used in RT-PCR, 40 of 133 samples (30.1%) tested positive for the presence of enterovirus RNA. When hepatitis A virus-specific primers were used, 12 of 139 samples (8.6%) were considered positive for the presence of hepatitis A viral RNA. The RT-PCR analysis performed with rotavirus-specific primers identified 18 of 130 samples (13.8%) that were positive for rotavirus RNA sequences. Our sample-processing technique and large-volume PCR protocol (reaction volume, 300 μl) resulted in sufficient removal or dilution of inhibitors so that more than 95% of the samples could be assayed by PCR. Because of its sensitivity for detecting viral nucleic acid sequences, PCR analysis should produce more positive results than cell culture analysis. Since either cell culture analysis or PCR can reveal only a “snapshot” of the quality of the groundwater being sampled, PCR seems to be a desirable rapid initial screening tool.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Reference15 articles.
1. Detection of enteroviruses in groundwater with the polymerase chain reaction
2. Craun
G. F.
Health aspects of groundwater pollution
Groundwater pollution microbiology.
Bitton
G.
Gerba
C. P.
1984
135
179
John Wiley & Sons Inc.
New York N.Y
3. DeLeon
R.
Shieh
C.
Baric
R. S.
Sobsey
M. D.
Detection of enteroviruses and hepatitis A virus in environmental samples by gene probes and polymerase chain reaction. Advances in water analysis and treatment
Proceedings of the 1990 American Water Works Association WQTC
1990
833
853
4. Edzwald
J. K.
Van Benschoten
J. E.
Aluminum coagulation of natural organic matter
Proceedings of the 4th Gothenburg Symposium on Chemical Water and Wastewater Treatment
1990
341
359
5. Polymerase chain reaction for the human picornaviruses.;Hyypia T.;J. Gen. Virol.,1989
Cited by
123 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献