Rapid detection of rabies virus by reverse transcription loop-mediated isothermal amplification.
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.
Boldbaatar
Bazartseren
B
Department of Veterinary Science, National Institute of Infectious Diseases, Tokyo 162-8640, Japan.
Inoue
Satoshi
S
Sugiura
Naoko
N
Noguchi
Akira
A
Orbina
Jun Ryan C
JR
Demetria
Catalino
C
Miranda
Mary Elizabeth
ME
Yamada
Akio
A
D000818:Animals; D001483:Base Sequence; D002371:Cat Diseases / Q000175:diagnosis / Q000821:virology; D002415:Cats; D017931:DNA Primers; D004283:Dog Diseases / Q000175:diagnosis / Q000821:virology; D004285:Dogs; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D011818:Rabies / Q000175:diagnosis* / Q000662:veterinary / Q000821:virology; D011820:Rabies virus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D012367:RNA, Viral; D012194:RNA-Directed DNA Polymerase