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

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

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

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.

2015

rotavirus, Diagnóstico, PCR, PCR em tempo real, NSP5, suínos

DOI: 10.1590/S0100-736X2015000100009

Pesquisa Veterinária Brasileira

Colégio Brasileiro de Patologia Animal (CBPA)

Veterinary medicine

EN, PT