Rapid detection of porcine kobuvirus in feces by reverse transcription loop-mediated isothermal amplification

Virol J. 2014 Apr 23:11:73. doi: 10.1186/1743-422X-11-73.

Abstract

Background: PKV is a new emerging pathogen detected in diarrhea pigs. At present, no more detection methods were reported except RT-PCR method. this study was to develop a fast diagnostic method based on the LAMP reaction for rapid detection of PKV nucleic acid in fecal samples.

Findings: Two pairs of primers were designed to amplify the conservative 3D gene of PKV genome. The PKV RT-LAMP method possessed well specificity and had 100 times higher sensitivity than common reverse transcription PCR (RT-PCR), which could detect up to 10 RNA copies of the target gene.

Conclusions: The results showed that the optimal reaction condition for RT-LAMP was achieved at 64°C for 50 min. Furthermore, the RT-LAMP procedure does not demand special equipment and is time-saving.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers / genetics
  • Feces / virology*
  • Kobuvirus / isolation & purification*
  • Nucleic Acid Amplification Techniques / methods*
  • Picornaviridae Infections / diagnosis
  • Picornaviridae Infections / veterinary*
  • Picornaviridae Infections / virology
  • RNA, Viral / genetics
  • Reverse Transcription*
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / diagnosis*
  • Swine Diseases / virology*
  • Temperature
  • Time

Substances

  • DNA Primers
  • RNA, Viral