Development of a Real-time PCR test for porcine group A rotavirus diagnosis

Pesquisa Veterinária Brasileira

Marconi, Elizabeth C.M.¹  Beserra, Laila A.R.¹  Bernardes, Nara T.C.G.¹  Silva, Fernanda D.F.¹  Gregori, Fabio¹ 

[1] Universidade de São Paulo, São Paulo, Brazil

关键词: Rotavirus;    diagnosis;    PCR;    Real-Time PCR;    NSP5;    swineRESUMORotavírus do grupo A (RVA) é uma das causas mais frequentes de diarreias em humanos e várias espécies animais;   

DOI  :  10.1590/S0100-736X2015000100009

来源: Colegio Brasileiro de Patologia Animal-CBPA

【 摘 要 】

Group A Rotavirus (RVA) is one of the most common causes of diarrhea in humans and several animal species. A SYBR-Green Real-Time polymerase chain reaction (PCR) was developed to diagnose RVA from porcine fecal samples, targeting amplification of a 137-bp fragment of nonstructural protein 5 (NSP5) gene using mRNA of bovine NADH-desidrogenase-5 as exogenous internal control. Sixty-five samples were tested (25 tested positive for conventional PCR and genetic sequencing). The overall agreement (kappa) was 0.843, indicating 'very good' concordance between tests, presenting 100% of relative sensitivity (25+ Real Time PCR/25+ Conventional PCR) and 87.5% of relative sensitivity (35- Real Time PCR/40- Conventional PCR). The results also demonstrated high intra- and inter-assay reproducibility (coefficient of variation ≤1.42%); thus, this method proved to be a fast and sensitive approach for the diagnosis of RVA in pigs.

【 授权许可】

Unknown