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

authors:

avatar Qian-Feng Xia 1 , avatar Yang-An Wen 2 , avatar Ping Liu 2 , avatar Pu Li 2 , avatar Jin-Bo Liu 2 , avatar Xi Qin 1 , avatar Shi-Yun Qian 1 , avatar Zhi-Guang Tu 3 , *

1) The Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, the Faculty of Laboratory Medicine, Chongqing Medical University,Chongqing. 2) The Faculty of Laboratory Medicine and Tropical Medicine, Hainan Medical College, China
The Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, the Faculty of Laboratory Medicine, Chongqing Medical University, China
The Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, the Faculty of Laboratory Medicine, Chongqing Medical University, zhiguangtu@sina.com, China
Hepatitis Monthly: Vol.11, issue 7; 519-524
article type: Research Article
received: December 6, 2010
accepted: April 1, 2011

how to cite: Xia Q, Wen Y, Liu P, Li P, Liu J, et al. Use of duplex mutation primers for real-time PCR quantification of hepatitis C virus RNA in serum. Hepat Mon. 2011;11(7): 519-524. 

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.

© 2011 Kowsar M.P.Co. All rights reserved.

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