Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region

Detalhes bibliográficos
Autor(a) principal: Nakatani, Sueli M.
Data de Publicação: 2010
Outros Autores: Santos, Carlos A., Riediger, Irina Nastassja, Krieger, Marco Aurélio, Duarte, Cesar A. B., Lacerda, Marco A., Biondo, Alexander W., Carrilho, Flair J., Ono-Nita, Suzane K.
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/39915
Resumo: Laboratório Central do Estado do Paraná. São José dos Pinhais, PR, Brasil / Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil.
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spelling Nakatani, Sueli M.Santos, Carlos A.Riediger, Irina NastassjaKrieger, Marco AurélioDuarte, Cesar A. B.Lacerda, Marco A.Biondo, Alexander W.Carrilho, Flair J.Ono-Nita, Suzane K.2020-02-12T19:56:58Z2020-02-12T19:56:58Z2010NAKATANI, Sueli M. et al. Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region. Plos One. v. 5, n. 4, p. 1-6, 2010.1932-6203https://www.arca.fiocruz.br/handle/icict/3991510.1371/journal.pone.00101501932-6203porPublic Library of ScienceHepatitis C virusPCRHepacivirusGenotyping TechniquesReal-Time Polymerase Chain ReactionTécnicas de GenotipajeReacción en Cadena en Tiempo Real de la PolimerasaTécnicas de GenotipagemReação em Cadeia da Polimerase em Tempo RealDevelopment of hepatitis C virus genotyping by real-time PCR based on the NS5B regioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleLaboratório Central do Estado do Paraná. São José dos Pinhais, PR, Brasil / Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil.Centro de Genomas. São Paulo, SP, Brasil.Laboratório Central do Estado do Paraná. São José dos Pinhais, PR, Brasil / Universidade Federal do Paraná. Laboratório de Biologia Celular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Division of Gastroenterology and Hepatology. Indiana University. Indianapolis, Indiana, USA.Universidade Federal do Paraná. Departamento de Medicina Veterinária. Curitiba, PR, Brasil / Department of Pathobiology. University of Illinois. Urbana, Illinois, USA.Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil.Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil.Hepatitis C virus (HCV) genotyping is the most significant predictor of the response to antiviral therapy. The aim of this study was to develop and evaluate a novel real-time PCR method for HCV genotyping based on the NS5B region. As methodology/principal findings: Two triplex reaction sets were designed, one to detect genotypes 1a, 1b and 3a; and another to detect genotypes 2a, 2b, and 2c. This approach had an overall sensitivity of 97.0%, detecting 295 of the 304 tested samples. All samples genotyped by real-time PCR had the same type that was assigned using LiPA version 1 (Line in Probe Assay). Although LiPA v. 1 was not able to subtype 68 of the 295 samples (23.0%) and rendered different subtype results from those assigned by real-time PCR for 12/295 samples (4.0%), NS5B sequencing and real-time PCR results agreed in all 146 tested cases. Analytical sensitivity of the real-time PCR assay was determined by end-point dilution of the 5000 IU/ ml member of the OptiQuant HCV RNA panel. The lower limit of detection was estimated to be 125 IU/ml for genotype 3a, 250 IU/ml for genotypes 1b and 2b, and 500 IU/ml for genotype 1a. And as significant conclusions had: The total time required for performing this assay was two hours, compared to four hours required for LiPA v. 1 after PCR-amplification. Furthermore, the estimated reaction cost was nine times lower than that of available commercial methods in Brazil. Thus, we have developed an efficient, feasible, and affordable method for HCV genotype identification.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/39915/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINALjournal.pone.0010150ok.PDFjournal.pone.0010150ok.PDFapplication/pdf219886https://www.arca.fiocruz.br/bitstream/icict/39915/2/journal.pone.0010150ok.PDFa79437d7da5af244a2756140499f6e86MD52TEXTjournal.pone.0010150ok.PDF.txtjournal.pone.0010150ok.PDF.txtExtracted texttext/plain34581https://www.arca.fiocruz.br/bitstream/icict/39915/3/journal.pone.0010150ok.PDF.txt4f56cb711a38a7adca2356c09ad5c796MD53icict/399152020-02-13 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dc.title.pt_BR.fl_str_mv Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
title Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
spellingShingle Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
Nakatani, Sueli M.
Hepatitis C virus
PCR
Hepacivirus
Genotyping Techniques
Real-Time Polymerase Chain Reaction
Técnicas de Genotipaje
Reacción en Cadena en Tiempo Real de la Polimerasa
Técnicas de Genotipagem
Reação em Cadeia da Polimerase em Tempo Real
title_short Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
title_full Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
title_fullStr Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
title_full_unstemmed Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
title_sort Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region
author Nakatani, Sueli M.
author_facet Nakatani, Sueli M.
Santos, Carlos A.
Riediger, Irina Nastassja
Krieger, Marco Aurélio
Duarte, Cesar A. B.
Lacerda, Marco A.
Biondo, Alexander W.
Carrilho, Flair J.
Ono-Nita, Suzane K.
author_role author
author2 Santos, Carlos A.
Riediger, Irina Nastassja
Krieger, Marco Aurélio
Duarte, Cesar A. B.
Lacerda, Marco A.
Biondo, Alexander W.
Carrilho, Flair J.
Ono-Nita, Suzane K.
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Nakatani, Sueli M.
Santos, Carlos A.
Riediger, Irina Nastassja
Krieger, Marco Aurélio
Duarte, Cesar A. B.
Lacerda, Marco A.
Biondo, Alexander W.
Carrilho, Flair J.
Ono-Nita, Suzane K.
dc.subject.other.pt_BR.fl_str_mv Hepatitis C virus
PCR
topic Hepatitis C virus
PCR
Hepacivirus
Genotyping Techniques
Real-Time Polymerase Chain Reaction
Técnicas de Genotipaje
Reacción en Cadena en Tiempo Real de la Polimerasa
Técnicas de Genotipagem
Reação em Cadeia da Polimerase em Tempo Real
dc.subject.en.pt_BR.fl_str_mv Hepacivirus
Genotyping Techniques
Real-Time Polymerase Chain Reaction
dc.subject.es.pt_BR.fl_str_mv Técnicas de Genotipaje
Reacción en Cadena en Tiempo Real de la Polimerasa
dc.subject.decs.pt_BR.fl_str_mv Técnicas de Genotipagem
Reação em Cadeia da Polimerase em Tempo Real
description Laboratório Central do Estado do Paraná. São José dos Pinhais, PR, Brasil / Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil.
publishDate 2010
dc.date.issued.fl_str_mv 2010
dc.date.accessioned.fl_str_mv 2020-02-12T19:56:58Z
dc.date.available.fl_str_mv 2020-02-12T19:56:58Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.citation.fl_str_mv NAKATANI, Sueli M. et al. Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region. Plos One. v. 5, n. 4, p. 1-6, 2010.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/39915
dc.identifier.issn.pt_BR.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0010150
dc.identifier.eissn.none.fl_str_mv 1932-6203
identifier_str_mv NAKATANI, Sueli M. et al. Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region. Plos One. v. 5, n. 4, p. 1-6, 2010.
1932-6203
10.1371/journal.pone.0010150
url https://www.arca.fiocruz.br/handle/icict/39915
dc.language.iso.fl_str_mv por
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
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