PMID: 8609195Nov 1, 1995Paper

Increased sensitivity of poliovirus detection in tap water concentrates by reverse transcriptase-polymerase chain reaction

Journal of Virological Methods
J F MaI L Pepper

Abstract

This study developed a methodology to increase the sensitivity of enteric virus detection in tap water concentrates. Polymerase chain reaction (PCR) detection of virus in reduced volumes of virus-containing water concentrates was successful following removal of PCR inhibitory substances. Poliovirus 1 and coxsackievirus B3 were seeded into 378 l of tap water, concentrated with 1MDS filters, and reconcentrated by organic flocculation. The volume of concentrates was successfully reduced from 25 to 5 ml without loss of virus recovery. PCR detection of virus after treatment of a water concentrate (1.1 x 10(5)-fold concentration) with a Sephadex G-100 plus Chelex-100 column, or Sephadex G-50 plus Chelex-100 column, followed by heat treatment to release viral RNA, was compared with direct phenol-chloroform-isoamyl alcohol (PCI) extraction of viral RNA. The Sephadex G-50 plus Chelex-100 column did not remove inhibitory substances efficiently. The Sephadex G-100 plus Chelex-100 column could remove inhibitory substances, however, 99% of the viruses were also removed by the column. PCI extraction was found to be sufficient to remove inhibitory substances for reverse transcriptase (RT)-seminested PCR with a sensitivity of 0.2 plaque-formin...Continue Reading

References

Oct 1, 1988·Journal of Virological Methods·K J RichardsonC P Gerba

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Citations

Mar 2, 2006·International Journal of Environmental Health Research·Stacey Yong Foong YeeYong Su Ming
Sep 30, 2000·Applied and Environmental Microbiology·P W HuangX Jiang
Feb 21, 2006·FEMS Immunology and Medical Microbiology·Tanxi CaiKehe Huang
Mar 23, 2001·Canadian Journal of Microbiology·K A ReynoldsL L Pepper
Aug 14, 2002·World Journal of Gastroenterology : WJG·Jun-Wen LiFu-Huan Chao
Apr 1, 1996·Applied and Environmental Microbiology·K A ReynoldsI L Pepper

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