Validation of single real-time TaqMan PCR assay for the detection and quantitation of four major genotypes of hepatitis E virus in clinical specimens

J Med Virol. 2006 Aug;78(8):1076-82. doi: 10.1002/jmv.20665.

Abstract

Since the characterization of the genome of the hepatitis E virus (HEV) in 1990, a large genetic diversity has been described. A single real-time reverse transcription (RT)-PCR assay with TaqMan technology has been validated which uses only one set of primers and probe within the ORF2 HEV region (nt 5207-5292) for the detection and quantification of the four major genotypes of HEV. This assay proved to be as efficient as the conventional RT-PCR methodology for the detection of HEV in clinical samples testing positive previously. The real-time RT-PCR and conventional RT-PCR were performed comparatively on 60 pairs of sera and stools collected during a recent outbreak of hepatitis E in Darfur. The real-time RT-PCR assay was 10- to 100-fold sensitive than for conventional RT-PCR assays used in this study with a range quantitation from 1.8 x 10(1) to 7.2 x 10(3) RNA copies/microl in clinical samples (serum and stools).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Genome, Viral
  • Genotype
  • Hepatitis E / virology*
  • Hepatitis E virus / classification
  • Hepatitis E virus / genetics*
  • Hepatitis E virus / isolation & purification*
  • Humans
  • Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Taq Polymerase / metabolism*

Substances

  • Taq Polymerase