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

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 impleme...

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Bibliographic Details
Published in:Memórias do Instituto Oswaldo Cruz 2016-06, Vol.111 (6), p.385-390
Main Authors: Tolardo, Aline Lavado, Souza, William Marciel de, Romeiro, Marilia Farignoli, Vieira, Luiz Carlos, Souza Luna, Luciano Kleber de, Henriques, Dyana Alves, Araujo, Jansen de, Hassegawa Siqueira, Carlos Eduardo, Colombo, Tatiana Elias, Aquino, Victor Hugo, Fonseca, Benedito Antonio Lopes da, Bronzoni, Roberta Vieira de Morais, Nogueira, Maurício Lacerda, Durigon, Edison Luiz, Figueiredo, Luiz Tadeu Moraes
Format: Article
Language:English
Subjects:
Animals
Cattle
diagnosis of vesicular stomatitis
Horses - virology
Humans
PARASITOLOGY
quantitative real-time RT-PCR
Real-Time Polymerase Chain Reaction
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
RNA, Viral - genetics
Sensitivity and Specificity
TROPICAL MEDICINE
Vesicular Stomatitis - diagnosis
Vesiculovirus
Vesiculovirus - genetics
zoonotic virus
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Summary: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.
ISSN:1678-8060
0074-0276
1678-8060
DOI:10.1590/0074-02760150456