title: Detection of avian group D rotavirus using the polymerase chain reaction for the VP6 gene.
abstract: 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/μL (0.5 pg/μL) 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.
authors: Seção de Virologia, Instituto Evandro Chagas, Secretaria de Vigilância em Saúde, Ministério da Saúde, Rodovia BR 316-KM 07, S/N, Levilândia, 67.030-000, Ananindeua, Pará, Brazil.
authors: Delana Andreza Melo
authors: DA
authors: Bezerra
authors: René Ribeiro
authors: RR
authors: da Silva
authors: Jane Haruko Lima
authors: JH
authors: Kaiano
authors: Rodrigo Vellasco Duarte
authors: RV
authors: Silvestre
authors: Darleise
authors: D
authors: de Souza Oliveira
authors: Alexandre C
authors: AC
authors: Linhares
authors: Yvone Benchimol
authors: YB
authors: Gabbay
authors: Joana D'Arc Pereira
authors: JD
authors: Mascarenhas
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics*; D001483:Base Sequence; D036022:Capsid Proteins / Q000032:analysis / Q000235:genetics*; D002645:Chickens / Q000821:virology; D003001:Cloning, Molecular; D017931:DNA Primers / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics; D005243:Feces / Q000821:virology; D005814:Genes, Viral; D010802:Phylogeny; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D017931:DNA Primers; D012367:RNA, Viral; C061332:VP6 protein, Rotavirus