title: In-house quantitative real-time PCR for the diagnosis of hepatitis B virus and hepatitis C virus infections.
abstract: The quantification of viral nucleic acids in serum by real-time PCR plays an important role in diagnosing hepatitis B virus and hepatitis C virus infection. In this study, we developed an assay using specific primers and probes to quantify hepatitis B virus DNA or hepatitis C virus RNA in serum from infected patients. For standardization and validation of the assay, an international panel of hepatitis B virus/hepatitis C virus and standard plasmids was used. A correlation coefficient of 0.983 and 0.963 for hepatitis B virus and hepatitis C virus, respectively, was obtained based on cycle threshold values and concentrations of DNA or RNA. The standard curve showed a linear relationship from 19IU/mL to 1.9×109IU/mL of serum, with a coefficient of determination (r2) of 0.99. In sera from patients infected with hepatitis B virus or hepatitis C virus viral loads (19IU/mL and 1.9×109IU/mL), we quantified viral loads with a detection limit of 1.9×102IU/mL. The real-time quantitative PCR assay developed in this study provides an ideal system for routine diagnosis and confirmation of indeterminate serological results, especially in immunosuppressed patients.
authors.affiliation: Instituto Hermes Pardini, Departamento de Pesquisa e Desenvolvimento, Vespasiano, MG, Brazil. Electronic address: danielle.zauli@hermespardini.com.br.
authors.forename: Danielle Alves Gomes
authors.initials: DA
authors.lastname: Zauli
authors.affiliation: Linhagen Produtos em Biotecnologia Ltda, Belo Horizonte, Minas Gerais, Brazil.
authors.forename: Carla Lisandre Paula de
authors.initials: CL
authors.lastname: Menezes
authors.affiliation: Instituto Hermes Pardini, Departamento de Pesquisa e Desenvolvimento, Vespasiano, MG, Brazil.
authors.forename: Cristiane Lommez de
authors.initials: CL
authors.lastname: Oliveira
authors.affiliation: Instituto Hermes Pardini, Departamento de Pesquisa e Desenvolvimento, Vespasiano, MG, Brazil.
authors.forename: Elvis Cristian Cueva
authors.initials: EC
authors.lastname: Mateo
authors.affiliation: Instituto Hermes Pardini, Departamento de Pesquisa e Desenvolvimento, Vespasiano, MG, Brazil.
authors.forename: Alessandro Clayton de Souza
authors.initials: AC
authors.lastname: Ferreira
mesh_terms: D004279:DNA, Viral; D016174:Hepacivirus / Q000235:genetics*; D006509:Hepatitis B / Q000175:diagnosis* / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics*; D006526:Hepatitis C / Q000175:diagnosis* / Q000821:virology*; D006801:Humans; D012367:RNA, Viral; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019562:Viral Load
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral
keywords: Hepatitis B; Hepatitis C; qPCR