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Updated: Aug 9, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Strategy for the maximization of clinically relevant information from hepatitis C virus, RT-PCR quantification
J Levis1, E Kenny-Walsh, K O'Sullivan
1Hepatitis C Unit, Department of Medicine, Clinical Sciences Building, Cork University Hospital, University College, Cork, Ireland.
Background:
The increasing clinical application of viral load assays for monitoring viral infections has been an incentive for the development of standardized tests for the hepatitis C virus.
Objective:
To develop a simple model for the prediction of baseline viral load in individuals infected with the hepatitis C virus.
Methodology:
Viral load quantification of each patient's first sample was assessed by RT-PCR-ELISA using the Roche MONITOR assay in triplicate. Genotype of the infecting virus was identified by reverse line probe hybridization, using amplicons resulting from the qualitative HCV Roche AMPLICOR assay.
Results:
Retrospective evaluation of first quantitative values suggested that 82.4% (n=168/204) of individuals had a viral load between 4.3 and 6.7 log(10) viral copies per ml. A few patients (3.4%; n=7/204) have a serum viremia less than the lower limit of the linear range of the RT-PCR assay. Subsequent, prospective evaluation of hepatitis C viral load of all new patients using a model based on the dynamic range of viral load in the retrospective group correctly predicted the dynamic range in 75.9% (n=33/54).
Conclusion:
The dynamic range of hepatitis C viremia extends beyond the linear range of the Roche MONITOR assay. Accurate determination of serum viremia is substantially improved by dilution of specimens prior to quantification.
