用聚合酶链反应单链构象多态性分析技术检测结核分枝杆菌L型的rpoB基因突变

Detecting rpoB gene mutation in Mycobacterium tuberculosis L-form isolated from pneu moconiosis patients by PCR-single-strand conformation polymorphism analysis

目的:探索结核分枝杆菌L型的耐药基因rpoB突变与利福平耐药性的关系,以及聚合酶链反应单链构象多态性分析(PCR-SSCP)技术的临床应用价值。方法:收集结核分枝杆菌L型菌株52株,采用PCR-SSCP方法检测rpoB基因,与采用常规药敏试验法检测利福平耐药的结果进行对比分析。结果:采用常规法检测,52株临床分离株中有38株利福平耐药,耐药率为73.08%(38/52);PCR-SSCP方法对52株临床耐药株rpoB基因突变的检出率为65.38%(32/52),两种方法检测耐利福平的符合率为84.21%。结论:PCR-SSCP对利福平耐药的结核分枝杆菌L型菌株检出率较高,将其与常规药敏试验联合应用,是一种指导临床用药的好方法。

Objective：To study the relation of rpoB gene mutation to resistance of rifampin in Mycobacterium tuberculosis L - form and explore PCR - single - strand conformation polymorphism analysis （SSCP） used in clinical application, nethods：A total of 52 clinical isolates of Mycobacterium tuberculosis L - form have been collected, and to compare the rpoB gene mutation detecting by PCR - SSCP with that conventional susceptibility testing. Results： There were 38 multi - resistance strains in the 52 isolates, which resistance - rate was 73.08% （38/52）, and gene mutation rate of rpoB was 65. 38% （32/52） by PCR - SSCP, coincidence rate of two methods was 84. 21% （32/38）. Conclusion：The combined application of PCR -SSCP analysis and conventional susceptibility test may be favourable to direct the therapeutic practice.