Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum

Abstract

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 (r2 = 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(r2 = 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.

Implication for health policy/practice/research/medical education:This novel HCV diagnostic assay is recommended to all clinicians involved in diagnosis and treatment of patients with HCV infection. We also suggest reader's attentions in the field of real-time PCR and liver infection diseases to the conclusion of this article. Please cite this paper as: Xia QF, Wen YA, Liu P, Li P, Liu JB, Qin X, et al. Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum. Hepat Mon. 2011;11(7):519-24.

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