Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/14940
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
Authors: Botelho-Souza, Luan Felipo
Santos, Alcione de Oliveira dos
Borzacov, Lourdes Maria
Honda, Eduardo Resende
Villalobos-Salcedo, Juan Miguel
Vieira, Deusilene Souza
Affilliation: 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.
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.
Abstract: 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.
Keywords: Hepatitis delta vírus
HDV RNA
Quantitative real-time PCR
Hydrolysis probe
Viral load
Issue Date: 2014
Publisher: Elsevier
Citation: 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.
DOI: 10.1016/j.jviromet.2013.11.016
ISSN: 0166-0934
Copyright: open access
Appears in Collections:RO - Artigos de Periódicos

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