Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock
Korean Journal of Veterinary Research
;
: 37-42, 2017.
Artigo
em Inglês
| WPRIM
| ID: wpr-91209
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Suínos
/
DNA
/
RNA
/
RNA Viral
/
Glicoproteínas
/
Reação em Cadeia da Polimerase
/
Surtos de Doenças
/
Sensibilidade e Especificidade
/
Genoma
/
Alphavirus
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Inglês
Revista:
Korean Journal of Veterinary Research
Ano de publicação:
2017
Tipo de documento:
Artigo
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