title: Comparison of conventional PCR with real-time PCR and branched DNA-based assays for hepatitis C virus RNA quantification and clinical significance for genotypes 1 to 5.
abstract: The key parameter for diagnosis and management of hepatitis C virus (HCV) infection is HCV RNA. Standardization of HCV RNA assays to IU is mainly based on genotype 1 panels. Little is known about the variability of commercially available HCV RNA assays for quantification of different genotypes. Two real-time reverse transcription (RT)-PCR assays (COBAS TaqMan HCV Test for use with the High-Pure System [HPS/CTM] and COBAS Ampliprep/COBAS TaqMan HCV Test [CAP/CTM]), one standard RT-PCR assay (COBAS Amplicor HCV Monitor 2.0 [CAM]), and one signal amplification assay (Versant Quantitative 3.0 [branched DNA [bDNA]]) were compared for quantification of genotypes 1 to 5 (n = 108). Using CAM as a reference assay for genotype 1-infected patients, the mean interassay differences compared with CAP/CTM, HPS/CTM, and bDNA were 0.16, -0.13, and -0.48 log(10) IU/ml HCV RNA, respectively. Comparison of CAM with CAP/CTM, HPS/CTM, and bDNA for the remaining genotypes showed the following results, respectively: 2a/c, -0.24, -0.78, and -0.49; 2b, -0.21, -0.18, and -0.64; 3a, 0.13, -1.04, and -0.55; 4, -0.52, -1.51, and -0.05; and 5, -0.28, -1.00, and -0.24 log IU/ml HCV RNA. A correct decision for treatment discontinuation in genotype 1 patients at week 12 was possible only when the same assay was used at baseline and week 12. Comparison of CAM with the CAP/CTM assay showed equal quantifications of genotype 1, 2, 3, and 5 samples, while genotype 4 samples were slightly underestimated. For the HPS/CTM assay, a significant underestimation of the HCV RNA concentrations of genotypes 2a/c, 3, 4, and 5 was observed. For the bDNA assay, a constant lower quantification of genotypes 1 to 3 was detected.
authors.affiliation: Klinik für Innere Medizin II, Universitätsklinikum des Saarlandes, Kirrberger Str., D-66421 Homburg/Saar, Germany. christoph.sarrazin@uniklinik-saarland.de
authors.forename: Christoph
authors.initials: C
authors.lastname: Sarrazin
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Gärtner
authors.forename: Dorothea
authors.initials: D
authors.lastname: Sizmann
authors.forename: Rainer
authors.initials: R
authors.lastname: Babiel
authors.forename: Ulrike
authors.initials: U
authors.lastname: Mihm
authors.forename: Wolf Peter
authors.initials: WP
authors.lastname: Hofmann
authors.forename: Michael
authors.initials: M
authors.lastname: von Wagner
authors.forename: Stefan
authors.initials: S
authors.lastname: Zeuzem
mesh_terms: D004279:DNA, Viral / Q000235:genetics; D005838:Genotype; D016174:Hepacivirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006526:Hepatitis C / Q000175:diagnosis / Q000188:drug therapy / Q000821:virology; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D011446:Prospective Studies; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral