Development of a New TaqMan-MGB Probe-Based Real-Time qPCR Method for the Detection of HCV-RNA
10.3969/j.issn.1671-7414.2017.05.009
- VernacularTitle:新型TaqMan-MGB探针实时荧光定量PCR检测HCV-RNA方法学的建立
- Author:
Bin QIAO
1
;
Yan LI
;
Ming WANG
;
Ge XIONG
Author Information
1. 武汉大学人民医院检验科
- Keywords:
TaqMan-MGB probe;
real-time qPCR;
HCV-RNA;
pseudoviral particles
- From:
Journal of Modern Laboratory Medicine
2017;32(5):32-35
- CountryChina
- Language:Chinese
-
Abstract:
Objective Developing a rapid and accurate real-time qPCR method for the detection of HCV-RNA.Methods HCV nucleotide sequence was analysed in Clustal software and primers and probe were designed in the conserved region of 5'UTR.The reaction system optimization of real-time qPCR method was used chessboard titration,pseudoviral particles were used as quantitative standard to assess the performance.New methods was compared with clinical commonly used kit of HCV-RNA and discuss the application value.Results The sensitivity of new real-time qPCR method was 50 IU/ml,coefficient variation was less than 5％.The quantitative results of this method could be traceable to national standards of GBW09151a.40 samples were determined by new methods and clinical commonly used kit of HCV-RNA,the positive concordance rate was 100 ％,the negative concordance rate was 56 ％.14 samples were positive by new method,but negative by Qiagen kit,illustrating that the sensitivity of new method was superior to Qiagen kit.Conclusion New TaqMan-MGB probe-based real-time qPCR method is a specific,sensitive,simple,rapid and exactly used to detection of HCV-RNA.
