Rapid Detection of Rabies Virus by Reverse Transcription Loop-Mediated Isothermal Amplification

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  • Boldbaatar Bazartseren
    Department of Veterinary Science, National Institute of Infectious Diseases, Japan The United Graduate School of Veterinary Science, Gifu University, Japan
  • Inoue Satoshi
    Department of Veterinary Science, National Institute of Infectious Diseases, Japan The United Graduate School of Veterinary Science, Gifu University, Japan
  • Sugiura Naoko
    Department of Veterinary Science, National Institute of Infectious Diseases, Japan The United Graduate School of Veterinary Science, Gifu University, Japan
  • Noguchi Akira
    The United Graduate School of Veterinary Science, Gifu University, Japan
  • Orbina Jun Ryan C.
    Department of Health, Research Institute for Tropical Medicine, the Philippines
  • Demetria Catalino
    Department of Health, Research Institute for Tropical Medicine, the Philippines
  • Miranda Mary Elizabeth
    Department of Health, Research Institute for Tropical Medicine, the Philippines
  • Yamada Akio
    Department of Veterinary Science, National Institute of Infectious Diseases, Japan The United Graduate School of Veterinary Science, Gifu University, Japan

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<p>In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was established which can detect 103 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.<tt> </tt></p>

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