Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction.
We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR.
Bonutti
Daniela Wey
DW
Centro de Pesquisa em Virologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Av. Bandeirantes 3900, 14049-000 Ribeirão Preto, SP, Brasil.
Figueiredo
Luiz Tadeu Moraes
LT
D000818:Animals; D001938:Brazil; D016679:Genome, Viral; D006023:Glycoproteins / Q000235:genetics*; D006168:Guinea Pigs; D051379:Mice; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018353:Rhabdoviridae Infections / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D018116:Vesiculovirus / Q000145:classification / Q000235:genetics*
D006023:Glycoproteins; D012367:RNA, Viral