Rapid detection of rabies virus by reverse transcription loop-mediated isothermal amplification

Jpn J Infect Dis. 2009 May;62(3):187-91.

Abstract

In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was established which can detect 10(3) copies of viral RNA corresponding to approximately 5 fg of RNA. RT-LAMP with the Phil primer set designed according to the nucleotide sequences obtained from a Kyoto patient who contracted rabies in the Philippines was able to amplify all 16 street viral sequences derived from the Philippines. The specificity of RT-LAMP products was easily confirmed by digestion with RsaI restriction enzyme. The reaction of RT-LAMP could be completed within 1 h and could be conducted under isothermal conditions using a conventional water bath or heat blocks, indicating that RT-LAMP is ideal for the diagnosis of rabies in developing countries. Although further study is required to establish more universal RT-LAMP primers applicable to viruses from other regions or countries, the fast, easy, simple, sensitive and specific RT-LAMP method established here might be useful for rabies diagnosis and can facilitate studies of rabies epidemiology where rabies is enzootic, particularly in developing countries.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cat Diseases / diagnosis
  • Cat Diseases / virology
  • Cats
  • DNA Primers
  • Dog Diseases / diagnosis
  • Dog Diseases / virology
  • Dogs
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Amplification Techniques / methods*
  • RNA, Viral / analysis
  • RNA, Viral / metabolism
  • RNA-Directed DNA Polymerase / metabolism*
  • Rabies / diagnosis*
  • Rabies / veterinary
  • Rabies / virology
  • Rabies virus / genetics
  • Rabies virus / isolation & purification*
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • RNA, Viral
  • RNA-Directed DNA Polymerase