Development of PCR methods for enteric virus detection in water

K. J. Schwab, R. De Leon, M. D. Sobsey

Research output: Contribution to journalConference article

Abstract

This study developed a reverse transcriptase-polymerase chain reaction (RT-PCR) detection system for enteric viruses in sample concentrates obtained by conventional filter adsorption-elution methods. One liter beef extract (BE)-glycine (G) eluant seeded with poliovirus 1 and hepatitis A virus (HAV) was used as a model system and the eluant further processed for RT-PCR compatibility. Sample concentration and purification procedures consisted of polyethylene glycol (PEG) precipitation, Pro-Cipitate (Affinity Technology, New Brunswick, NJ) precipitation, a second PEG precipitation, spin-column chromatography, and ultrafiltration. Sample volumes are reduced from 1 L to 20-40 μL and purified sufficiently for viral detection by RT-PCR. As little as 3 PFU of poliovirus 1 in an initial 1 L eluate were detected by RT-PCR.

Original languageEnglish (US)
Pages (from-to)211-218
Number of pages8
JournalWater Science and Technology
Volume27
Issue number3-4
DOIs
StatePublished - Jan 1 1993
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

  • Beef extract
  • Concentration
  • Enteric viruses
  • PEG precipitation
  • Purification
  • Rt-PCR
  • Spin-column chromatography

ASJC Scopus subject areas

  • Environmental Engineering
  • Water Science and Technology

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