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

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.