title: Quantitation of replication of the HCV genome in human livers with end-stage cirrhosis by strand-specific real-time RT-PCR assays: methods and clinical relevance.
abstract: HCV replicates in liver via an intermediate negative strand RNA. To study the relevance of HCV genome replication, quantitative strand-specific HCV real-time RT-PCR assays were developed and applied to livers explanted because of end-stage cirrhosis. The assays have broad ranges of determination and a high reproducibility and accuracy. Analysis of five different samples showed an even distribution of HCV genomes in four livers. Hepatic concentrations of positive (PS)- and negative (NS)-strand RNA did correlate with each other, with PS/NS ratios ranging between 3 and 340. Hepatic concentrations of HCV-PS or -NS RNA did not correlate with serum HCV-RNA levels or with genotypes. A high HCV envelope-2 protein expression correlated with a low NS concentration. HCV-PS and -NS levels, E2 protein expression and genotype did not correlate with biochemical tests or with histological changes in the explanted liver, but the ratio NS/PS, a marker of viral replication, correlated with the severity of the recurrent post-transplant hepatitis caused by HCV. This suggests the existence of an extra-hepatic location of HCV with comparable viral replication rate being responsible for the infection of the newly transplanted liver.
authors.affiliation: Department of Hepatology, University Hospitals, Leuven, Belgium.
authors.forename: Lan
authors.initials: L
authors.lastname: Lin
authors.forename: Louis
authors.initials: L
authors.lastname: Libbrecht
authors.forename: Jannick
authors.initials: J
authors.lastname: Verbeeck
authors.forename: Chris
authors.initials: C
authors.lastname: Verslype
authors.forename: Tania
authors.initials: T
authors.lastname: Roskams
authors.forename: Jos
authors.initials: J
authors.lastname: van Pelt
authors.forename: Marc
authors.initials: M
authors.lastname: Van Ranst
authors.forename: Johan
authors.initials: J
authors.lastname: Fevery
mesh_terms: D000328:Adult; D000368:Aged; D005260:Female; D016174:Hepacivirus / Q000254:growth & development*; D019698:Hepatitis C, Chronic / Q000150:complications* / Q000821:virology*; D006801:Humans; D008103:Liver Cirrhosis / Q000821:virology*; D008297:Male; D008875:Middle Aged; D012367:RNA, Viral / Q000096:biosynthesis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; D014764:Viral Proteins