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KMID : 0359320170570010037
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Korean Journal of Veterinary Research
2017 Volume.57 No. 1 p.37 ~ p.42
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Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock
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Lee Seung-Heon
Yang Dong-Kun
Kim Ha-Hyun
Choi Sung-Suk
Cho In-Soo
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Abstract
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Getah virus (GETV) infection causes sporadic outbreaks of mild febrile illness in horses and reproductive failure in pigs. In this study, we established a reverse transcription polymerase chain reaction (RT-PCR) method to detect GETV from suspected virus-infected samples. The reaction conditions were optimized and validated by using RNA extracted from GETV propagated in cell culture. A GETV-specific GED4 primer set was designed and used to amplify a 177 bp DNA fragment from a highly conserved region of the E1 glycoprotein gene in the GETV genome. RT-PCR performed with this primer set revealed high sensitivity and specificity. In the sensitivity test, the GED4 primer set detected GETV RNA at the level of 102.0 TCID50/mL. In the specificity test, the GED4 primer set amplified only a single band of PCR product on the GETV RNA template, without non-specific amplification, and exhibited no crossreactivity with other viral RNAs. These results suggest that this newly established RT-PCR method is useful for accurate identification of GETV infection in animals.
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KEYWORD
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Getah virus, diagnosis, reverse transcription polymerase chain reaction
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