A novel method for liberating viral nucleic acid for assay of water samples with cDNA probes

Kenneth J. Richardson, Aaron B. Margolin, Charles P. Gerba

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Rapid and sensitive methods are needed for the detection of enteric viruses to ensure proper drinking water quality. Gene probes have been shown to be useful for this purpose. Previously, samples to be assayed were treated with a series of phenol-chloroform extractions to release the viral nucleic acid. We have developed a more rapid procedure for liberating or exposing the genome of poliovirus for probing. In this study, a poliovirus model was used to test the ability of heat (65°C for 30 min) for release or exposure of viral nucleic acid. Several different RNase inhibitors were tested for their ability to prevent viral RNA degradation. A comparison of the two methods indicates phenol-chloroform extraction is not necessary before probing. In addition to saving 2-4 h of time, maximum sensitivity levels were consistently obtained using this novel procedure.

Original languageEnglish (US)
Pages (from-to)13-21
Number of pages9
JournalJournal of Virological Methods
Volume22
Issue number1
DOIs
StatePublished - Oct 1988

Keywords

  • Dot-blot hybridization
  • Heat treatment
  • Human placental RNasin
  • Phenol-chloroform extraction
  • Poliovirus

ASJC Scopus subject areas

  • Virology

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