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Updated: Aug 13, 2026
A Protocol for Analyzing Hepatitis C Virus Replication
Published on: June 26, 2014
Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction
M Abdel-Hamid1, D C Edelman, W E Highsmith
1University of Maryland School of Medicine, Department of Pathology, Baltimore, USA.
Objectives:
To compare the performance of reverse transcription followed by the polymerase chain reaction (RT-PCR), without RNA purification, with the performance of classic protocols.
Study Design/Methods:
Direct and classic techniques were used to test three groups of samples: six hepatitis C virus (HCV) seroconversion panels (n = 90), a HCV RNA reference panel (n = 26), and serial dilutions of four HCV-positive sera (n = 24). These methods were then applied sequentially through a clinical diagnostic algorithm to test 268 samples from high-risk patients.
Results:
For the three groups of samples, we found a 94% concordance between direct and purified RT-PCR methods. For the detection of HCV RNA in clinical samples, sensitivity was maximized and cost minimized using both protocols according to the proposed algorithm.
Conclusions:
The direct PCR method is reliable, sensitive, and can result in time and cost savings. The suggested testing algorithm can enhance sensitivity and time savings for populations with a high prevalence of infection.
