Development of duplex RT-PCR-ELISA for the simultaneous detection of hepatitis A virus and hepatitis E virus

J Virol Methods. 2011 Jul;175(1):137-40. doi: 10.1016/j.jviromet.2011.04.030. Epub 2011 May 4.

Abstract

This study aimed to develop a specific and sensitive duplex reverse transcription polymerase chain reaction enzyme-linked immunosorbent assay (duplex RT-PCR-ELISA) for hepatitis A virus (HAV) and hepatitis E virus (HEV). Duplex RT-PCR-ELISA could detect and differentiate HAV and HEV with specific probes. When ELISA technique was used to detect probe-bound RT-PCR products, duplex RT-PCR-ELISA could detect as little as 0.1 ng/μL HAV and HEV from clinical samples. Human norovirus, enterovirus, poliovirus, murine norovirus and feline calicivirus were used for the specificity test; all were negative. Therefore duplex RT-PCR-ELISA can be used for the simultaneous detection of HAV and HEV in contaminated fecal samples.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme-Linked Immunosorbent Assay / methods*
  • Hepatitis A / virology*
  • Hepatitis A virus / genetics
  • Hepatitis A virus / isolation & purification*
  • Hepatitis E / virology*
  • Hepatitis E virus / genetics
  • Hepatitis E virus / isolation & purification*
  • Humans
  • Real-Time Polymerase Chain Reaction / methods*