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. 2011 Jul;11(7):519-24.

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

Qian-Feng Xia  1 , Yang-An Wen, Ping Liu, Pu Li, Jin-Bo Liu, Xi Qin, Shi-Yun Qian, Zhi-Guang Tu
Affiliations

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

Qian-Feng Xia et al. Hepat Mon. 2011 Jul.

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 (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.

Keywords: Duplex mutation primers; HCV; Hepatitis; PCR; Quantification.

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Conflict of interest statement

Conflict of interest: None declared.

Figures

Figure 1
Figure 1
Fluorescent curves of the standard dilution series.
Figure 2
Figure 2
Graph shin correlation of the HCV real-time PCR ass av results with the new method results for 92 HCV-positive clinical spcimens (numerical values e not listed). Thre was excellent correlation (r 0. 95) between the two assays for all samples.
See this image and copyright information in PMC

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