Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction
We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR.
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Instituto Oswaldo Cruz, Ministério da Saúde
2005
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oai:scielo:S0074-027620050002000142005-07-22Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reactionBonutti,Daniela WeyFigueiredo,Luiz Tadeu Moraes Vesiculovirus vesicular stomatitis virus reverse transcription-polymerase chain reaction diagnosis nested-PCR Brazil We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR.info:eu-repo/semantics/openAccessInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz v.100 n.2 20052005-04-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762005000200014en10.1590/S0074-02762005000200014 |
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Bonutti,Daniela Wey Figueiredo,Luiz Tadeu Moraes |
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Bonutti,Daniela Wey Figueiredo,Luiz Tadeu Moraes Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
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Bonutti,Daniela Wey Figueiredo,Luiz Tadeu Moraes |
author_sort |
Bonutti,Daniela Wey |
title |
Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
title_short |
Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
title_full |
Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
title_fullStr |
Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
title_full_unstemmed |
Diagnosis of Brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
title_sort |
diagnosis of brazilian vesiculoviruses by reverse transcription-polymerase chain reaction |
description |
We describe a reverse transcription-polymerase chain reaction (RT-PCR) and a nested-PCR for diagnosis of Piry, Carajás, Cocal, and Alagoas vesiculoviruses from Brazil. The RNA extracts of viral and clinical samples were submitted to a RT-PCR using Vesiculovirus G primers that amplify part of the glycoprotein gene. The RT-PCR produced amplicons of expected size, 290 base pair, for the four studied viruses. The RT-PCR showed a high sensitivity being 151.3 times (2.18 log) more sensitive for the detection of Piry virus than the classical procedure for virus detection in tissue culture based on the viral cytophatic effect. Amplicons had nucleotides sequenced and were aligned in order to select internal primers for a nested-PCR to confirm the origin of Piry, Carajás, Cocal, and Alagoas Vesiculovirus. Ten blood and tarsal pad epithelial samples of infected Guinea-pigs had Vesiculovirus genome amplified by RT-nested-PCR. |
publisher |
Instituto Oswaldo Cruz, Ministério da Saúde |
publishDate |
2005 |
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http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762005000200014 |
work_keys_str_mv |
AT bonuttidanielawey diagnosisofbrazilianvesiculovirusesbyreversetranscriptionpolymerasechainreaction AT figueiredoluiztadeumoraes diagnosisofbrazilianvesiculovirusesbyreversetranscriptionpolymerasechainreaction |
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