title: 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.
  
  
  OBJECTIVE
  To develop and validate a novel real-time PCR for quantification of HCV RNA based on the duplex mutation primers technology.
  
  
  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.
authors.affiliation: The Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, the Faculty of Laboratory Medicine, Chongqing Medical University, Chongqing, China.
authors.forename: Qian-Feng
authors.initials: QF
authors.lastname: Xia
authors.forename: Yang-An
authors.initials: YA
authors.lastname: Wen
authors.forename: Ping
authors.initials: P
authors.lastname: Liu
authors.forename: Pu
authors.initials: P
authors.lastname: Li
authors.forename: Jin-Bo
authors.initials: JB
authors.lastname: Liu
authors.forename: Xi
authors.initials: X
authors.lastname: Qin
authors.forename: Shi-Yun
authors.initials: SY
authors.lastname: Qian
authors.forename: Zhi-Guang
authors.initials: ZG
authors.lastname: Tu
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