Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction

Memórias do Instituto Oswaldo Cruz

Daniela Wey Bonutti¹  Luiz Tadeu Moraes Figueiredo¹ 

[1] ,Universidade de São Paulo Faculdade de Medicina de Ribeirão Preto Centro de Pesquisa em VirologiaRibeirão Preto SP ,Brasil

关键词: Vesiculovirus;    vesicular stomatitis virus;    reverse transcription-polymerase chain reaction;    diagnosis;    nested-PCR;    Brazil;   

DOI  :  10.1590/S0074-02762005000200014

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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