荧光PCR探针熔解曲线法检测结核分枝杆菌耐利福平突变研究

    马艳艳1，李辉2，赵东阳2，李静1，李庆阁3

    1.河南国际旅行卫生保健中心，河南 郑州450016；2.河南省疾病预防控制中心;

    3.厦门大学生命科学学院生物医学科学系

    摘要：目的对荧光PCR熔解曲线法检测结核分枝杆菌耐利福平突变方法进行临床研究，评价检测能力及在国境口岸的应用价值。方法应用荧光PCR熔解曲线法检测结核病人临床分离结核分枝杆菌，以传统药物敏感性试验为标准，获得该方法的灵敏性、特异性。结果对347例结核病人临床分离培养样本，传统药物敏感性试验检出271例利福平敏感标本，76例利福平耐药标本。荧光PCR熔解曲线法检出269例利福平敏感标本，78例利福平耐药标本，灵敏性为93.42%，特异性为97.42%，符合率为96.54%。结论荧光PCR熔解曲线法检测速度快速、灵敏度高、特异性强，可用于结核分枝杆菌利福平耐药突变的快速检测，适于国境口岸对耐多药结核病的快速筛查。

    关键词：结核分枝杆菌；熔解曲线法；利福平；检测

    中图分类号：R52　文献标识码：B

    Evaluation of the mutations detection of rpoBgene in

    Mycobacterium tuberculosis clinical isolatesby probe

    melting analysis based real-time PCR

    MA Yan-yan， LI hui， ZHAO Dong-yang， LIJing，LI Qing-ge

    Henan International Travel Health CareCenter, Zhengzhou, Heanan 450016,China

    Abstract:   Objective  To evaluate the analysis abilities and valuation of the meltinganalysis based real-time PCR for detecting the mutations of ropBgene in mycobacterium tuberculosis from clinical samples atfrontier port.  Methods   The probe melting analysisassay was adopted for the sensitivity and specificity in detectingthe isolates from clinical samples against the standard of drugsusceptibility testing (DST).   Results   Atotal of 347 mycobacterium tuberculosis isolates were detected bytwo methods, in which, 271 rifampin sensitivity isolates and 76rifampin resistant isolates were gained by DST. Meanwhile, 269rifampin sensitivity isolates and 78 rifampin resistant isolateswere gained by the probe melting analysis assay, the sensitivityand specificity of melting analysis assay was 93.42% and 97.42%,respectively. The concordance of the two methods was 96.54%. Conclusion   The probe melting analysis assay own thecharacteristics of short-time, higher sensitivity and specificityfor detecting rifampin resistance in tuberculosis using clinicalsample and is available to use for rapid screening multidrug drugresistant tuberculosis at frontier ports.

    Key words:   Mycobacteriumtuberculosis; Melting analysis assay; Rifampin; Detect

    《中国国境卫生检疫杂志》