Development of a quantitative real-time RT-PCR assay for detecting Taiwan ferret badger rabies virus in ear tissue of ferret badgers and mice
Journal
Journal of Veterinary Medical Science
Journal Volume
80
Journal Issue
6
Pages
1012-1019
Date Issued
2018
Author(s)
Hsu, Ai-Ping
Tseng, Chun-Hsien
Lu, Yi-Ta
Shih, Yu-Hua
Chen, Re-Shang
Tsai, Kuo-Jung
Tu, Wen-Jane
Cliquet, Florence
Abstract
In 2013, the first case of Taiwan ferret badger rabies virus (RABV-TWFB) infection was reported in Formosan ferret badgers, and two genetic groups of the virus were distinguished through phylogenetic analysis. To detect RABV-TWFB using a sensitive nucleic acid-based method, a quantitative real-time reverse transcription polymerase chain reaction targeting the conserved region of both genetic groups of RABV-TWFB was developed. This method had a limit of detection (LOD) of 40 RNA copies/reaction and detected viral RNA in brain and ear tissue specimens of infected and dead Formosan ferret badgers and mice with 100% sensitivity and specificity. The mean viral RNA load detected in the ear tissue specimens of ferret badgers ranged from 3.89 × 108 to 9.73 × 108 RNA copies/g-organ, which was 111-fold to 2,220-fold lower than the concentration detected in the brain specimens, but 2,000-fold to 5,000-fold higher than the LOD of the assay. This highly sensitive technique does not require facilities or instruments complying with strict biosafety criteria. Furthermore, it is efficient, safe, and labor-saving as only ear specimens need be sampled. Therefore, it is a promising technique for epidemiological screening of Taiwan ferret badger rabies. © 2018 The Japanese Society of Veterinary Science.
Subjects
Rabies diagnosis; Real-time RT-PCR; Taiwan ferret badger rabies
SDGs
Other Subjects
animal; Bagg albino mouse; isolation and purification; mouse; Mustela putorius furo; Mustelidae; phylogeny; rabies; Rabies virus; reverse transcription polymerase chain reaction; Taiwan; veterinary medicine; virology
Type
journal article