Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil

Detalhes bibliográficos
Autor(a) principal: Botelho-Souza, Luan Felipo
Data de Publicação: 2014
Outros Autores: Santos, Alcione de Oliveira dos, Borzacov, Lourdes Maria, Honda, Eduardo Resende, Villalobos-Salcedo, Juan Miguel, Vieira, Deusilene Souza
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/14940
Resumo: Fundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.
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spelling Botelho-Souza, Luan FelipoSantos, Alcione de Oliveira dosBorzacov, Lourdes MariaHonda, Eduardo ResendeVillalobos-Salcedo, Juan MiguelVieira, Deusilene Souza2016-07-14T15:34:41Z2016-07-15T18:40:34Z2016-07-15T18:40:34Z2014BOTELHO-SOUZA, Luan Felipo et al. Development of a reverse transcription quantitative real-timePCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil. Journal of Virological Methods, v. 197, p. 19-24, 2014.0166-0934https://www.arca.fiocruz.br/handle/icict/1494010.1016/j.jviromet.2013.11.016engElsevierDevelopment of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazilinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil.Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.Fundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.The hepatitis delta virus (HDV) is a pathogen that causes a severe and rapidly progressive disease ofhepatocytes. The measurement of viral load in the peripheral blood of patients with HDV infections isimportant for diagnosis, treatment monitoring, and support for follow-up studies of viral replication dur-ing the course of the disease. This study reports the development of an assay capable of detecting andquantifying the abundance of HDV particles in serum samples, based on reverse-transcription quantita-tive PCR (RT-qPCR). Two standards for calibration were produced for determining the viral load of HDV:a cDNA cloned into a linear plasmid and a transcribed RNA. For validating this assay, 140 clinical samplesof sera were used, comprising 100 samples from patients who tested positive for anti-HDV and hepatitisB virus surface antigen (HBsAg) by ELISA; 30 samples from blood donors; 5 samples monoinfected withhepatitis B virus (HBV); and 5 samples monoinfected with hepatitis C virus (HCV). The HDV RT-qPCR assayperformed better when calibrated using the standard based on HDV cDNA cloned into a linear plasmid,yielding an efficiency of 99.8% and a specificity of 100% in the in vitro assays. This study represents thefirst HDV RT-qPCR assay developed with clinical samples from Brazil and offers great potential for newclinical efficacy studies of antiviral therapeutics for use in patients with hepatitis delta in the western Amazon region.Hepatitis delta vírusHDV RNAQuantitative real-time PCRHydrolysis probeViral loadinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZTEXTDevelopment of a reverse transcription.pdf.txtDevelopment of a reverse transcription.pdf.txtExtracted texttext/plain35171https://www.arca.fiocruz.br/bitstream/icict/14940/4/Development%20of%20a%20reverse%20transcription.pdf.txt66954954a32e342f185d534363acf5f8MD54ORIGINALDevelopment of a reverse 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dc.title.pt_BR.fl_str_mv Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
title Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
spellingShingle Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
Botelho-Souza, Luan Felipo
Hepatitis delta vírus
HDV RNA
Quantitative real-time PCR
Hydrolysis probe
Viral load
title_short Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
title_full Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
title_fullStr Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
title_full_unstemmed Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
title_sort Development of a reverse transcription quantitative real-time PCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil
author Botelho-Souza, Luan Felipo
author_facet Botelho-Souza, Luan Felipo
Santos, Alcione de Oliveira dos
Borzacov, Lourdes Maria
Honda, Eduardo Resende
Villalobos-Salcedo, Juan Miguel
Vieira, Deusilene Souza
author_role author
author2 Santos, Alcione de Oliveira dos
Borzacov, Lourdes Maria
Honda, Eduardo Resende
Villalobos-Salcedo, Juan Miguel
Vieira, Deusilene Souza
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Botelho-Souza, Luan Felipo
Santos, Alcione de Oliveira dos
Borzacov, Lourdes Maria
Honda, Eduardo Resende
Villalobos-Salcedo, Juan Miguel
Vieira, Deusilene Souza
dc.subject.en.eng.fl_str_mv Hepatitis delta vírus
HDV RNA
Quantitative real-time PCR
Hydrolysis probe
Viral load
topic Hepatitis delta vírus
HDV RNA
Quantitative real-time PCR
Hydrolysis probe
Viral load
description Fundação Oswaldo Cruz. Fiocruz Rondônia. Laboratório Plataforma Técnica. Porto Velho, RO, Brazil / Centro de Pesquisa em Medicina Tropical de Rondônia. Porto Velho, RO, Brazil / Universidade Federal de Rondônia. Departamento de Medicina. Núcleo de Saúde. Programa de Pós-graduação em Biologia Experimental. Porto Velho, RO, Brazil.
publishDate 2014
dc.date.issued.fl_str_mv 2014
dc.date.accessioned.fl_str_mv 2016-07-14T15:34:41Z
2016-07-15T18:40:34Z
dc.date.available.fl_str_mv 2016-07-15T18:40:34Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv BOTELHO-SOUZA, Luan Felipo et al. Development of a reverse transcription quantitative real-timePCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil. Journal of Virological Methods, v. 197, p. 19-24, 2014.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/14940
dc.identifier.issn.pt_BR.fl_str_mv 0166-0934
dc.identifier.doi.pt_BR.fl_str_mv 10.1016/j.jviromet.2013.11.016
identifier_str_mv BOTELHO-SOUZA, Luan Felipo et al. Development of a reverse transcription quantitative real-timePCR-based system for rapid detection and quantitation of hepatitis delta virus in the western Amazon region of Brazil. Journal of Virological Methods, v. 197, p. 19-24, 2014.
0166-0934
10.1016/j.jviromet.2013.11.016
url https://www.arca.fiocruz.br/handle/icict/14940
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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