Comparison of cell culture and a poliovirus gene probe assay for the detection of enteroviruses in environmental water samples

A. B. Margolin, C. P. Gerba, K. J. Richardson, J. E. Naranjo

Research output: Contribution to journalConference articlepeer-review

7 Scopus citations

Abstract

Nucleic acid hybridization provides a rapid non-cell culture method for the detection of enteric viruses in water. The purpose of this work was to compare the detection of naturally occurring enteroviruses by cell culture with their detection by a poliovirus gene probe in various types of water samples. Samples of activated sludge effluent, tertiary treated wastewater (activated sludge, filtration and passage through reverse osmosis), ground water, surface water and tidal river water were processed through 1 MDS Virozorb filters to concentrate any naturally occurring virus. Viruses were eluted from the filters with pH 9.5 beef extract and reduced in volume by flocculation to 20-30 ml. These concentrates were then assayed in the BGM cell line by the cytophatogenic effects (CPE) method and by a poliovirus cDNA probe (base pairs 115-7440) labeled with 32P. A total of 233 samples were assayed in this manner. In slightly more than 93% of the samples gene probe and cell culture yielded the same results. Of these samples 36 were positive by gene probe and 28 by cell culture assay. Positive samples for gene probe were confirmed by treatment with NaOH or RNAse and then reprobed. Samples demonstrating CPE upon primary passage were confirmed positive by subsequent passage of cell lysate on a new monolayer of BGM cells. Ten samples were positive by gene probe and negative by cell culture, and 4 samples were negative by gene probe and positive by cell culture.

Original languageEnglish (US)
Pages (from-to)311-314
Number of pages4
JournalWater Science and Technology
Volume27
Issue number3-4
DOIs
StatePublished - Jan 1 1993
Externally publishedYes
EventProceedings of the 16th Biennial Conference and Exposition of the International Association on Water Pollution Research and Control - Washington, DC, USA
Duration: May 24 1992May 30 1992

Keywords

  • Cell culture
  • Enteric viruses
  • Gene probes
  • Water
  • cDNA

ASJC Scopus subject areas

  • Environmental Engineering
  • Water Science and Technology

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