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Development and application of a real-time RT-PCR for brazilian Vesiculovirus

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Author(s):
Aline Lavado Tolardo
Total Authors: 1
Document type: Master's Dissertation
Press: Ribeirão Preto.
Institution: Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC)
Defense date:
Examining board members:
Luiz Tadeu Moraes Figueiredo; Eurico de Arruda Neto; Aramis Augusto Pinto
Advisor: Luiz Tadeu Moraes Figueiredo
Abstract

The Vesiculovirus is a Rhabdoviridae family genre of RNA virus that includes serotypes Carajás, Cocal, Maraba, Piry, Alagoas and Indiana. These are causes of vesicular stomatitis in ruminants and human febrile illness in Brazil. The vesiculoviroses and its epidemiology are little known in humans. Still, Vesiculovirus (VSV) are poorly diagnosed in humans and laboratory animals by the lack of diagnostic methods. Therefore, we proposed in this work to develop and test a real-time RT-PCR by SYBR Green method, focusing on the detection of Brazilian VSV. Primers that amplify part of the VSV G protein gene were used in the test which proved capable of detecting genomes of VSV Piry, Indiana, Alagoas and Carajás. The method was used to test serum samples from patients with acute febrile disease, cattle, horses and macerated arthropods. Real time RT-PCR showed to be 100 times more sensitive than conventional RT-PCR for Vesiculovirus and also was possible to detect up to 10 RNA copies of the Piry virus. Also, the real-time RT-PCR for Vesiculovirus proved able to diagnose and quantify Alagoas VSV in serum samples from cattle and horses. Therefore, the real-time RT-PCR developed in this work will probably be very useful in the diagnosis and in future research, which will increase the epidemiological knowledge, as it is still little known about the Vesiculovirus. (AU)

FAPESP's process: 13/02256-2 - Development of Real Time RT-PCR for diagnosis of infections by Piry virus and other vesiculovirus
Grantee:Aline Lavado Tolardo
Support Opportunities: Scholarships in Brazil - Master