Quantitative Analysis of Serum Hepatitis C Virus RNA Levels by Computer-Assisted Analysis of Reverse-Transcription PCR Profiles

Abstract

Amplification of hepatitis C virus RNA by a two-step reverse-transcription (RT) and polymerase-chain-reaction (PCR) protocol is a well-established procedure for screening recently and chronically infected hepatitis C virus (HCV) patients. Quantification of HCV viremia is often done by more expensive or laborious procedures. In this study we present an alternative protocol for the semi-quantitative estimation of serum HCV RNA levels, based on a computer-based analysis of the electrophoresis profiles of amplified DNA obtained by RT-PCR. A non-linear regression calibration curve was established, based upon the results of a previous quantification assay for 13 chronically infected HCV patients. Quantitative analysis of serum RNA levels in 10 patients was evaluated on the basis of this calibration curve, indicating the correlation between the amount of the DNA in the electrophoresis profile and the number of viral copies per ml of serum.