title: An in-house real-time polymerase chain reaction: standardisation and comparison with the Cobas Amplicor HBV monitor and Cobas AmpliPrep/Cobas TaqMan HBV tests for the quantification of hepatitis B virus DNA.
abstract: This study aimed to standardise an in-house real-time polymerase chain reaction (rtPCR) to allow quantification of hepatitis B virus (HBV) DNA in serum or plasma samples, and to compare this method with two commercial assays, the Cobas Amplicor HBV monitor and the Cobas AmpliPrep/Cobas TaqMan HBV test. Samples from 397 patients from the state of São Paulo were analysed by all three methods. Fifty-two samples were from patients who were human immunodeficiency virus and hepatitis C virus positive, but HBV negative. Genotypes were characterised, and the viral load was measure in each sample. The in-house rtPCR showed an excellent success rate compared with commercial tests; inter-assay and intra-assay coefficients correlated with commercial tests (r = 0.96 and r = 0.913, p < 0.001) and the in-house test showed no genotype-dependent differences in detection and quantification rates. The in-house assay tested in this study could be used for screening and quantifying HBV DNA in order to monitor patients during therapy.
authors.affiliation: Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: Ana Paula de Torres
authors.initials: AP
authors.lastname: Santos
authors.affiliation: Laboratório de Virologia, Instituto de Medicina Tropical, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: José Eduardo
authors.initials: JE
authors.lastname: Levi
authors.affiliation: Núcleo de Doenças Sanguíneas e Sexuais, Centro de Virologia, Instituto Adolfo Lutz, São Paulo, SP, Brasil.
authors.forename: Marcilio Figueiredo
authors.initials: MF
authors.lastname: Lemos
authors.affiliation: Núcleo de Doenças Sanguíneas e Sexuais, Centro de Virologia, Instituto Adolfo Lutz, São Paulo, SP, Brasil.
authors.forename: Samira Julien
authors.initials: SJ
authors.lastname: Calux
authors.affiliation: Núcleo de Doenças Sanguíneas e Sexuais, Centro de Virologia, Instituto Adolfo Lutz, São Paulo, SP, Brasil.
authors.forename: Isabel Takano
authors.initials: IT
authors.lastname: Oba
authors.affiliation: Núcleo de Doenças Sanguíneas e Sexuais, Centro de Virologia, Instituto Adolfo Lutz, São Paulo, SP, Brasil.
authors.forename: Regina Célia
authors.initials: RC
authors.lastname: Moreira
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000592:standards; D004279:DNA, Viral / Q000302:isolation & purification*; D005069:Evaluation Studies as Topic; D005260:Female; D005838:Genotype; D060005:Genotyping Techniques / Q000592:standards*; D006679:HIV Seropositivity / Q000097:blood / Q000175:diagnosis; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000097:blood / Q000175:diagnosis*; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis; D006801:Humans; D007223:Infant; D062069:Inventions / Q000592:standards; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000295:instrumentation / Q000379:methods; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012680:Sensitivity and Specificity; D019562:Viral Load; D055815:Young Adult
chemical_list: D017931:DNA Primers; D004279:DNA, Viral