Detection of avian group D rotavirus using the polymerase chain reaction for VP6 gene

• Autor(a) principal: Bezerra, Delana Andreza Melo
• Data de Publicação: 2012
• Outros Autores: Silva, René Ribeiro da, Kaiano, Jane Haruko Lima, Silvestre, Rodrigo Vellasco Duarte, Oliveira, Darleise de Souza, Linhares, Alexandre da Costa, Gabbay, Yvone Benchimol, Mascarenhas, Joana D'Arc Pereira
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Digital do Instituto Evandro Chagas (Patuá)
• Texto Completo: https://patua.iec.gov.br/handle/iec/3269
• topic: Rotavirus; Infecções por Rotavirus; Diarreia; Aves; Reação em Cadeia da Polimerase Via Transcriptase Reversa

Group D rotaviruses (RVs-D) have been documented in birds and, while they may be common in these animals, few molecular studies are available for this specific group. In this study, specific primers for the gene that encodes for the RVs-D VP6 protein were designed and used in a reverse transcription polymerase chain reaction (RT-PCR). Thirty pools of samples were tested by polyacrylamide gel electrophoresis (PAGE) yielding a 30% (9/30) positivity. These pools were subjected subsequently to RT-PCR, with a 53% (16/30) positivity rate. The sensitivity of the PCR assay was demonstrated up to a dilution of 5 × 10−4 ng/L (0.5 pg/L) of the cloned VP6 gene. The four samples were sequenced and showed 90.8–91.1% similarity with regards to the RVs-D VP6 gene. To assess for specificity our RT-PCR was applied to nine samples known to contain enteric viral agents other than group D rotaviruses including picobirnavirus, rotavirus group A, and reovirus with negative results. Overall, the data confirm the specificity of the primers used for detecting the RVs-D by RT-PCR, suggesting that this assay can be used for diagnostic purposes.