Detection of enteroviruses in groundwater with the polymerase chain reaction

Morteza Abbaszadegan, M. S. Huber, C. P. Gerba, I. L. Pepper

Research output: Contribution to journalArticle

194 Citations (Scopus)

Abstract

Standard methods for the detection of enteroviruses in environmental samples involve the use of cell culture, which is expensive and time- consuming. The polymerase chain reaction (PCR) is an attractive method for the detection of enteroviruses in water because primary cell culture is not needed and the increased sensitivity of PCR allows detection of the low numbers of target DNAs and RNAs usually found in environmental samples. However, environmental samples often contain substances that inhibit PCR amplification of target DNA and RNA. Procedures that remove substances that interfere with the amplification process need to be developed if PCR is to be successfully applied to environmental samples. An RNA-PCR assay for the detection of enteroviruses in water was developed and used to test a variety of groundwater concentrates and humic acid solutions seeded with poliovirus type 1. The groundwater samples and humic acid solutions were treated with Sephadex G-50, Sephadex G-100, Sephadex G-200, Chelex-100 resin, and a mixed bed resin to remove PCR-inhibitory material from the samples. Sephadex G-100 in combination with Chelex-100 was found to be very effective in removing inhibitory factors for the detection of enteroviruses in groundwater concentrates by PCR. Viruses were detected in two of the groundwater concentrates by the RNA-PCR assay after treatment with Sephadex G-100 plus Chelex-100. This was confirmed by tissue culture, suggesting that the treatment protocol and, subsequently, the RNA-PCR assay are applicable for the detection of enteroviruses in environmental samples.

Original languageEnglish (US)
Pages (from-to)1318-1324
Number of pages7
JournalApplied and Environmental Microbiology
Volume59
Issue number5
StatePublished - 1993
Externally publishedYes

Fingerprint

Enterovirus
Groundwater
polymerase chain reaction
groundwater
Polymerase Chain Reaction
RNA
DNA-Directed RNA Polymerases
DNA-directed RNA polymerase
Humic Substances
concentrates
sampling
assay
humic acids
humic acid
resins
resin
amplification
cell culture
assays
enterovirus

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science(all)
  • Microbiology

Cite this

Detection of enteroviruses in groundwater with the polymerase chain reaction. / Abbaszadegan, Morteza; Huber, M. S.; Gerba, C. P.; Pepper, I. L.

In: Applied and Environmental Microbiology, Vol. 59, No. 5, 1993, p. 1318-1324.

Research output: Contribution to journalArticle

Abbaszadegan, Morteza ; Huber, M. S. ; Gerba, C. P. ; Pepper, I. L. / Detection of enteroviruses in groundwater with the polymerase chain reaction. In: Applied and Environmental Microbiology. 1993 ; Vol. 59, No. 5. pp. 1318-1324.
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