Korean J Vet Res.  2017 Mar;57(1):37-42. 10.14405/kjvr.2017.57.1.37.

Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock

• Lee SH
• Yang DK
• Kim HH
• Choi SS
• Cho IS

• KMID: 2376877
• DOI: http://doi.org/10.14405/kjvr.2017.57.1.37

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) TCIDâ‚…â‚€/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.

Keyword

Getah virus; diagnosis; reverse transcription polymerase chain reaction

MeSH Terms

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
DNA
Glycoproteins
RNA
RNA, Viral