title: A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay.
abstract: At present, the sporadic occurrence of human rabies in Brazil can be attributed primarily to dog- and vampire bat-related rabies viruses. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was employed as a simultaneous detection method for both rabies field variants within 60 min. Vampire bat-related rabies viruses could be distinguished from dog variants by digesting amplicons of the RT-LAMP reaction using the restriction enzyme AlwI. Amplification and digestion could both be completed within 120 min after RNA extraction. In addition, the RT-LAMP assay also detected rabies virus in isolates from Brazilian frugivorous bats and Ugandan dog, bovine and goat samples. In contrast, there were false negative results from several Brazilian insectivorous bats and all of Chinese dog, pig, and bovine samples using the RT-LAMP assay. This study showed that the RT-LAMP assay is effective for the rapid detection of rabies virus isolates from the primary reservoir in Brazil. Further improvements are necessary so that the RT-LAMP assay can be employed for the universal detection of genetic variants of rabies virus in the field.
authors.affiliation: Nihon University Veterinary Research Center, Fujisawa, Kanagawa 252-0880, Japan.
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Saitou
authors.forename: Yuki
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Shinji
authors.initials: S
authors.lastname: Hirano
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Mochizuki
authors.forename: Takuya
authors.initials: T
authors.lastname: Itou
authors.forename: Fumio H
authors.initials: FH
authors.lastname: Ito
authors.forename: Takeo
authors.initials: T
authors.lastname: Sakai
mesh_terms: D000818:Animals; D001938:Brazil; D002417:Cattle; D002685:Chiroptera; D017931:DNA Primers / Q000235:genetics; D004285:Dogs; D006041:Goats; D006801:Humans; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D011818:Rabies / Q000175:diagnosis / Q000662:veterinary*; D011820:Rabies virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D013552:Swine; D013997:Time Factors; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral