Use of Duplex Mutation Primers for Real-Time PCR Quantification of Hepatitis C Virus RNA in Serum
The duplex mutation primers offer many advantages over other multi-labeled probes for real-time detection of amplification products.
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.
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).
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.
The article is published online in Hepatitis Monthly and is free to access.