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Updated: May 27, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum
Qian-Feng Xia1, Yang-An Wen, Ping Liu
1The Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, the Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
Background:
The duplex mutation primers offer many advantages over other multi-labeled probes for real-time detection of amplification products.
Objectives:
To develop and validate a novel real-time PCR for quantification of HCV RNA based on the duplex mutation primers technology.
Materials And Methods:
The duplex mutation primers were selected in the highly conservative 5' non-coding region (5'NCR) of the HCV RNA. The assay was validated with the Viral Quality Control panel, which also includes Chinese HCV RNA standards.
Results:
The detection limit was 57 IU/mL, and a good linear correlation in the range of 102-108 IU/mL was revealed (r(2) = 0.999) with the novel method. This assay has a dynamic range of at least 8 log10 without the need for specimen dilution, good clinical intra- and inter-run precision, and excellent correlation with a commercially available assay(r(2) = 0.95).
Conclusions:
The high sensitivity, wide linear range, and good reproducibility, combined with low cost, make this novel quantitative HCV real-time PCR assay particularly well suited for application to clinical and epidemiological studies.
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