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

Aline Lavado Tolardo; 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

doi:10.1590/0074-02760150456

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

Aline Lavado Tolardo, 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

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

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 ± 0.8°C, 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.

Original language	English (US)
Pages (from-to)	385-390
Number of pages	6
Journal	Memorias do Instituto Oswaldo Cruz
Volume	111
Issue number	6
DOIs	https://doi.org/10.1590/0074-02760150456
State	Published - Jun 2016
Externally published	Yes

Keywords

Diagnosis of vesicular stomatitis
Quantitative real-time RT-PCR
Vesiculovirus
Zoonotic virus

ASJC Scopus subject areas

Microbiology (medical)

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Tolardo, A. L., de Souza, W. M., Romeiro, M. F., Vieira, L. C., Luna, L. K. D. S., Henriques, D. A., de Araujo, J., Siqueira, C. E. H., Colombo, T. E., Aquino, V. H., da Fonseca, B. A. L., Bronzoni, R. V. D. M., Nogueira, M. L., Durigon, E. L., & Figueiredo, L. T. M. (2016). A real-time reverse transcriptase polymerase chain reaction for detection and quantification of vesiculovirus. Memorias do Instituto Oswaldo Cruz, 111(6), 385-390. https://doi.org/10.1590/0074-02760150456

Tolardo, AL, de Souza, WM, Romeiro, MF, Vieira, LC, Luna, LKDS, Henriques, DA, de Araujo, J, Siqueira, CEH, Colombo, TE, Aquino, VH, da Fonseca, BAL, Bronzoni, RVDM, Nogueira, ML, Durigon, EL & Figueiredo, LTM 2016, 'A real-time reverse transcriptase polymerase chain reaction for detection and quantification of vesiculovirus', Memorias do Instituto Oswaldo Cruz, vol. 111, no. 6, pp. 385-390. https://doi.org/10.1590/0074-02760150456

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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 ± 0.8°C, 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.",

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AU - Luna, Luciano Kleber de Souza

AU - Henriques, Dyana Alves

AU - de Araujo, Jansen

AU - Siqueira, Carlos Eduardo Hassegawa

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AU - Aquino, Victor Hugo

AU - da Fonseca, Benedito Antonio Lopes

AU - Bronzoni, Roberta Vieira de Morais

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AU - Durigon, Edison Luiz

AU - Figueiredo, Luiz Tadeu Moraes

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N2 - 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 ± 0.8°C, 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.

AB - 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 ± 0.8°C, 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.

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

Abstract

Keywords

ASJC Scopus subject areas

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