Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
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Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock

  • Lee, Seung Heon (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Yang, Dong-Kun (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Kim, Ha-Hyun (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Choi, Sung-Suk (Viral Disease Division, Animal and Plant Quarantine Agency) ;
  • Cho, In-Soo (Viral Disease Division, Animal and Plant Quarantine Agency)
  • Received : 2017.02.15
  • Accepted : 2017.03.16
  • Published : 2017.03.31
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Abstract

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 $10^{2.0}\;TCID_{50}/mL$. 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 cross-reactivity 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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