Development of a New TaqMan-MGB Probe-Based Real-Time qPCR Method for the Detection of HCV-RNA / 现代检验医学杂志

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.