Abstract
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.
Highlights
Aline Lavado Tolardo1/+, William Marciel de Souza1, Marilia Farignoli Romeiro1, Luiz Carlos Vieira1, Luciano Kleber de Souza Luna1, Dyana Alves Henriques2, Jansen de Araujo2, Carlos Eduardo Hassegawa Siqueira3, Tatiana Elias Colombo4, Victor Hugo Aquino5, Benedito Antonio Lopes da Fonseca1, Roberta Vieira de Morais Bronzoni3, Maurício Lacerda Nogueira4, Edison Luiz Durigon2, Luiz Tadeu Moraes Figueiredo1
The real-time reverse transcriptase polymerase chain reaction (RT-PCR) showed a sensitivity of 10 copies/mL for all the VSV tested (Alagoas, Carajas, Piry vesiculovirus (PIRYV) and vesicular stomatitis Indiana virus (VSINV)) and specific TM peaks without the formation of primer-dimer for all of them, as shown in Table I and Fig. 2
We decided to develop a real-time RTPCR assay to detect a broad range of VSV species that could be used for epidemiological surveillance
Summary
Aline Lavado Tolardo1/+, William Marciel de Souza, Marilia Farignoli Romeiro, Luiz Carlos Vieira, Luciano Kleber de Souza Luna, Dyana Alves Henriques, Jansen de Araujo, Carlos Eduardo Hassegawa Siqueira, Tatiana Elias Colombo, Victor Hugo Aquino, Benedito Antonio Lopes da Fonseca, Roberta Vieira de Morais Bronzoni, Maurício Lacerda Nogueira, Edison Luiz Durigon, Luiz Tadeu Moraes Figueiredo. 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. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods This assay allows a preliminary differential diagnosis of VSV infections. From June-August 2013, cases of vesicular stomatitis disease affecting horses and cattle were reported to the Veterinary Hospital of the Federal University of Campina Grande, Paraiba state, Brazil. These outbreaks occurred in the towns of Paulista, São Bento, Patos, in Paraiba state, and Umarizal, Rio Grande do Norte state.
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