title: Usefulness of in-house PCR methods for hepatitis B virus DNA detection.
abstract: The aim of the present study was to evaluate the performance of three in-house PCR techniques for HBV DNA detection and compare it with commercial quantitative methods to evaluate the usefulness of in-house methods for HBV diagnosis. Three panels of HBsAg reactive sera samples were evaluated: (i) 50 samples were examined using three methods for in-house qualitative PCR and the Cobas Amplicor HBV Monitor Assay; (ii) 87 samples were assayed using in-house semi-nested PCR and the Cobas TaqMan HBV test; (iii) 11 serial samples obtained from 2 HBV-infected individuals were assayed using the Cobas Amplicor HBV test and semi-nested PCR. In panel I, HBV DNA was detected in 44 samples using the Cobas Amplicor HBV test, 42 samples using semi-nested PCR (90% concordance with Cobas Amplicor), 22 samples using PCR for the core gene (63.6% concordance) and 29 samples using single-round PCR for the pre-S/S gene (75% concordance). In panel II, HBV DNA was quantified in 78 of the 87 HBsAg reactive samples using Cobas TaqMan but 52 samples using semi-nested PCR (67.8% concordance). HBV DNA was detected in serial samples until the 17th and 26th week after first donation using in-house semi-nested PCR and the Cobas Amplicor HBV test, respectively. In-house semi-nested PCR presented adequate concordance with commercial methods as an alternative method for HBV molecular diagnosis in low-resource settings.
authors.affiliation: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Moyra Machado
authors.initials: MM
authors.lastname: Portilho
authors.affiliation: Laboratory of Enteroviruses, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Marcia Leite
authors.initials: ML
authors.lastname: Baptista
authors.affiliation: Department of Technical and Technological Support, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Messias
authors.initials: M
authors.lastname: da Silva
authors.affiliation: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Paulo Sérgio Fonseca
authors.initials: PS
authors.lastname: de Sousa
authors.affiliation: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Lia Laura
authors.initials: LL
authors.lastname: Lewis-Ximenez
authors.affiliation: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Lampe
authors.affiliation: Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil. Electronic address: liviafiocruz@gmail.com.
authors.forename: Livia Melo
authors.initials: LM
authors.lastname: Villar
mesh_terms: D000328:Adult; D000368:Aged; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D005260:Female; D006509:Hepatitis B / Q000175:diagnosis* / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D004279:DNA, Viral
keywords: Commercial methods; HBV DNA; In-house PCR; Molecular diagnosis