检验医学与临床2017，Vol.14Issue(11)：1528-1530,3.DOI:10.3969/j.issn.1672-9455.2017.11.003
实时荧光PCR检测结核杆菌DNA的分析灵敏度验证及临床应用
Analytic sensitivity verification and clinical application of real time fluorescent PCR for detecting Mycobacterium tuberculosis DNA
摘要
Abstract
Objective To establish and evaluate the real time fluorescent PCR qualitative method for analyzing the sensitivity verification method with Mycobacterium tuberculosis DNA(TB-DNA) detection as the entry point.Methods Five low concentration samples were taken and performed the parallel detection for 16 times,then the detection rate of each sample was calculated.The curve fitting was respectively performed by the concentrations logarithm(base-10),Ct value and detection probability.when the fluorescent PCR detection rate of TB-DNA was 50% and 95%,the corresponding concentration and Ct value were obtained,then the results report strategy was accordingly established.Fifty-two clinical samples were simultaneously detected by nest PCR and fluorescent PCR,the applicability of established results report strategy was evaluated.Results When TB-DNA detection rates were 50% and 95% respectively,the corresponding concentrations were 69 copy/mL and 2.46×103 copy/mL respectively.The established results report strategy of TB-DNA was report positive if the results >2.46×103 copy/mL,conducting re-detection if the results <2.46×103 copy/mL and >69 copy/mL,report positive if re-detection was positive,and report negative if the re-detection was negative,and report negative if the results <69 copy/mL.After detection in 52 clinical samples,6 cases were positive by fluorescent PCR,4 cases were positive by nest PCR.Calculating the Kappa value and u test(kappa=0.78,u=3.58,P<0.05) indicated that the results by the two kinds of method had good consistency.Conclusion Formulating the results report strategy of TB-DNA fluorescent PCR qualitative method according to the detection probability-concentration curve has good clinical applicability.Formulating the results report strategy according to the lowest detection limit can be used for the qualitative PCR based on the quantitative data.关键词
结核杆菌/最低检出限/方法学性能Key words
mycobacterium tuberculosis/lower detection limit/methodological performance引用本文复制引用
张鸿娟,宋宇,郭翀,赵滢,刘子杰..实时荧光PCR检测结核杆菌DNA的分析灵敏度验证及临床应用[J].检验医学与临床,2017,14(11):1528-1530,3.基金项目
云南省卫生系统高层次领军人才项目(L-2012-2). （L-2012-2）
