A method for simultaneous detection and identification of Brazilian dog- and vampire bat-related rabies virus by reverse transcription loop-mediated isothermal amplification assay

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 mi...

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Bibliographic Details
Published in:Journal of virological methods 2010-09, Vol.168 (1), p.13-17
Main Authors: Saitou, Yasumasa, Kobayashi, Yuki, Hirano, Shinji, Mochizuki, Nobuyuki, Itou, Takuya, Ito, Fumio H., Sakai, Takeo
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Brazil
Cattle
Chiroptera
Digestion
DNA Primers - genetics
Dog
Dogs
Fundamental and applied biological sciences. Psychology
Goats
Humans
Microbiology
Molecular Sequence Data
Nucleic Acid Amplification Techniques - methods
Rabies
Rabies - diagnosis
Rabies - veterinary
Rabies virus
Rabies virus - classification
Rabies virus - genetics
Rabies virus - isolation & purification
Reverse Transcription
RNA, Viral - genetics
RT-LAMP
Sensitivity and Specificity
Sequence Analysis, DNA
Swine
Techniques used in virology
Time Factors
Vampire bat
Virology
Virology - methods
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Summary: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.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2010.04.008