| Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock |
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Seung Heon Lee, Dong-Kun Yang, Ha-Hyun Kim, Sung-Suk Choi, In-Soo Cho |
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Viral Disease Division, Animal and Plant Quarantine Agency |
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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 $10^{2.0};TCID_{50}/mL$. 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. |
| Key Words:
Getah virus, diagnosis, reverse transcription polymerase chain reaction |
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