A strategy for detection of viruses in groundwater by PCR

Morteza Abbaszadegan, Peter Stewart, Mark LeChevallier

Research output: Contribution to journalArticle

140 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)444-449
Number of pages6
JournalApplied and Environmental Microbiology
Volume65
Issue number2
StatePublished - Feb 1999
Externally publishedYes

Fingerprint

Groundwater
virus
groundwater
Enterovirus
Viruses
Polymerase Chain Reaction
viruses
hepatitis
RNA
water industry
Rotavirus
sampling
Hepatitis A virus
cell culture
Water
Cell Culture Techniques
reverse transcriptase polymerase chain reaction
Industry
nucleic acid
water

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

A strategy for detection of viruses in groundwater by PCR. / Abbaszadegan, Morteza; Stewart, Peter; LeChevallier, Mark.

In: Applied and Environmental Microbiology, Vol. 65, No. 2, 02.1999, p. 444-449.

Research output: Contribution to journalArticle

Abbaszadegan, Morteza ; Stewart, Peter ; LeChevallier, Mark. / A strategy for detection of viruses in groundwater by PCR. In: Applied and Environmental Microbiology. 1999 ; Vol. 65, No. 2. pp. 444-449.
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