现代检验医学杂志2017，Vol.32Issue(5)：32-35,4.DOI:10.3969/j.issn.1671-7414.2017.05.009
新型TaqMan-MGB探针实时荧光定量PCR检测HCV-RNA方法学的建立
Development of a New TaqMan-MGB Probe-Based Real-Time qPCR Method for the Detection of HCV-RNA
乔斌 1李艳 1汪明 1熊格1
作者信息
- 1. 武汉大学人民医院检验科,武汉430060
- 折叠
摘要
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.关键词
TaqMan-MGB探针/实时荧光定量PCR/丙型肝炎病毒核酸/假病毒颗粒Key words
TaqMan-MGB probe/real-time qPCR/HCV-RNA/pseudoviral particles分类
生物科学引用本文复制引用
乔斌,李艳,汪明,熊格..新型TaqMan-MGB探针实时荧光定量PCR检测HCV-RNA方法学的建立[J].现代检验医学杂志,2017,32(5):32-35,4.
