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

• Autor(a) principal: Marconi,Elizabeth C.M.
• Data de Publicação: 2015
• Outros Autores: Bernardes,Nara T.C.G., Beserra,Laila A.R., Silva,Fernanda D.F., Gregori,Fabio
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Pesquisa Veterinária Brasileira (Online)
• Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2015000100039
• topic: Rotavirus; diagnosis; PCR; Real-Time PCR; NSP5; swine

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.