Detection and quantitation of hepatitis E virus in human faeces by real-time quantitative PCR

J Virol Methods. 2004 Jun 15;118(2):77-82. doi: 10.1016/j.jviromet.2004.01.025.

Abstract

Hepatitis E Virus (HEV) is the causative agent of an acute and self-limited form of hepatitis. The virus is transmitted by the faecal-oral route and is a major cause of viral hepatitis in much of the developing world where it causes sporadic infections and large-scale epidemics. A simple and rapid protocol for the measurement of HEV faecal shedding by a real-time polymerase chain reaction (PCR) with the SYBR Green method on a LightCycler instrument, is described. After only 3h the real-time quantitative PCR method detected 10 molecules of HEV cDNA fragment per reaction tube and showed a high linear dynamic range of quantitation (10-10(6) molecules of cDNA/reaction) with a good correlation (r = -1.00). Its specificity was confirmed by assay in human faecal samples.

Publication types

  • Evaluation Study

MeSH terms

  • Benzothiazoles
  • DNA, Complementary / genetics
  • Diamines
  • Feces / virology*
  • Hepatitis E / virology*
  • Hepatitis E virus / genetics*
  • Hepatitis E virus / isolation & purification*
  • Humans
  • Organic Chemicals / metabolism
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Quinolines
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity

Substances

  • Benzothiazoles
  • DNA, Complementary
  • Diamines
  • Organic Chemicals
  • Quinolines
  • SYBR Green I