Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock.
10.14405/kjvr.2017.57.1.37
- Author:
Seung Heon LEE
1
;
Dong Kun YANG
;
Ha Hyun KIM
;
Sung Suk CHOI
;
In Soo CHO
Author Information
1. Viral Disease Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Korea. yangdk@korea.kr
- Publication Type:Original Article
- Keywords:
Getah virus;
diagnosis;
reverse transcription polymerase chain reaction
- MeSH:
Alphavirus*;
Animals;
Cell Culture Techniques;
Diagnosis;
Disease Outbreaks;
DNA;
Genome;
Glycoproteins;
Horses;
Livestock*;
Methods;
Polymerase Chain Reaction*;
Reverse Transcription*;
RNA;
RNA, Viral;
Sensitivity and Specificity;
Swine
- From:Korean Journal of Veterinary Research
2017;57(1):37-42
- CountryRepublic of Korea
- Language:English
-
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) 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 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.
