Simple and rapid detection of swine hepatitis E virus by reverse transcription loop-mediated isothermal amplification

Arch Virol. 2012 Dec;157(12):2383-8. doi: 10.1007/s00705-012-1425-5. Epub 2012 Aug 2.

Abstract

Hepatitis E virus (HEV) is an enteric pathogen of humans and animals, and pigs have been considered an important reservoir of this virus. Recent evidence has indicated the cross-species transmission of hepatitis E virus (HEV) from pigs to humans, causing zoonosis, mostly via consumption of uncooked or undercooked animal meat/viscera. In this study, we have developed a one-step RT-LAMP assay for rapid detection of swine HEV. Specific primer sets targeting the ORF3 gene were designed. The sensitivity of the RT-LAMP assay was 10(1) copies/μl of RNA template, which was tenfold higher than that of RT-nPCR. The specificity of this assay was demonstrated by the lack of amplification of DNA/RNA from other swine viruses. Furthermore, a total of 41 bile samples were subjected to RT-LAMP and RT-nPCR. Eighteen positive samples were detected by RT-nPCR, while 36 positive samples were detected by RT-LAMP, indicating that the sensitivity of the RT-LAMP assay was higher than that of the conventional RT-nPCR assay. The RT-LAMP assay reported here may be used for diagnosis of swine HEV, not only in laboratories but also under field conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bile / virology
  • Gene Expression Regulation, Viral
  • Hepatitis E / diagnosis
  • Hepatitis E / veterinary*
  • Hepatitis E / virology
  • Hepatitis E virus / genetics
  • Hepatitis E virus / isolation & purification*
  • Nucleic Acid Amplification Techniques / methods
  • Nucleic Acid Amplification Techniques / veterinary*
  • Phylogeny
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis
  • Swine Diseases / virology*

Substances

  • RNA, Viral