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A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus

2016; Instituto Oswaldo Cruz, Ministério da Saúde; Volume: 111; Issue: 6 Linguagem: Inglês

10.1590/0074-02760150456

1678-8060

Aline Lavado Tolardo, William Marciel de Souza, Marília Farignoli Romeiro, Luiz Carlos Vieira, Luciano Kleber de Souza Luna, Dyana Alves Henriques, Jansen de Araújo, Carlos Eduardo Hassegawa Siqueira, Tatiana Elias Colombo, Víctor Hugo Aquino, Benedito Antônio Lopes da Fonseca, Roberta Vieira de Morais Bronzoni, Maurício Lacerda Nogueira, Edison Luíz Durigon, Luiz Tadeu Moraes Figueiredo,

Viral Infections and Immunology Research

Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.