| Author | Nakatani, Sueli M. | |
| Author | Santos, Carlos A. | |
| Author | Riediger, Irina Nastassja | |
| Author | Krieger, Marco Aurélio | |
| Author | Duarte, Cesar A. B. | |
| Author | Lacerda, Marco A. | |
| Author | Biondo, Alexander W. | |
| Author | Carrilho, Flair J. | |
| Author | Ono-Nita, Suzane K. | |
| Access date | 2020-02-12T19:56:58Z | |
| Available date | 2020-02-12T19:56:58Z | |
| Document date | 2010 | |
| Citation | 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. | pt_BR |
| ISSN | 1932-6203 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/39915 | |
| Language | por | pt_BR |
| Publisher | Public Library of Science | pt_BR |
| Rights | open access | pt_BR |
| Subject in Portuguese | Hepatitis C virus | pt_BR |
| Subject in Portuguese | PCR | pt_BR |
| Title | Development of hepatitis C virus genotyping by real-time PCR based on the NS5B region | pt_BR |
| Type | Article | pt_BR |
| DOI | 10.1371/journal.pone.0010150 | |
| Abstract | 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. | pt_BR |
| Affilliation | 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. | pt_BR |
| Affilliation | Centro de Genomas. São Paulo, SP, Brasil. | pt_BR |
| Affilliation | Laboratório Central do Estado do Paraná. São José dos Pinhais, PR, Brasil / Universidade Federal do Paraná. Laboratório de Biologia Celular. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil. | pt_BR |
| Affilliation | Division of Gastroenterology and Hepatology. Indiana University. Indianapolis, Indiana, USA. | pt_BR |
| Affilliation | Universidade Federal do Paraná. Departamento de Medicina Veterinária. Curitiba, PR, Brasil / Department of Pathobiology. University of Illinois. Urbana, Illinois, USA. | pt_BR |
| Affilliation | Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil. | pt_BR |
| Affilliation | Universidade de São Paulo. Faculdade de Medicina. Departamento de Gastroenterologia. São Paulo, SP, Brasil. | pt_BR |
| Subject | Hepacivirus | pt_BR |
| Subject | Genotyping Techniques | pt_BR |
| Subject | Real-Time Polymerase Chain Reaction | pt_BR |
| Subject in Spanish | Técnicas de Genotipaje | pt_BR |
| Subject in Spanish | Reacción en Cadena en Tiempo Real de la Polimerasa | pt_BR |
| DeCS | Técnicas de Genotipagem | pt_BR |
| DeCS | Reação em Cadeia da Polimerase em Tempo Real | pt_BR |
| e-ISSN | 1932-6203 | |
