快速检测结核分支杆菌对利福平的耐药性

Rapid detection of mycobacterium tuberculosis resistance to rifampin by moleccular method

目的 :建立快速检测结核分支杆菌耐药基因型的分子药敏试验方法。方法 :用聚合酶链反应 -单链构象多态性 (PCR -SSCP)分析结核分支杆菌rpoB基因突变并与药敏试验对照。结果 :116株结核分支杆菌的PCR -SSCP图谱与H37Rv为对照 ,81株耐药菌73株有明显区别 ,而35株敏感 ,仅1株位置有区别 ;检测阳性率90 1 % ;特异性97 1 %。结论 :rpoB基因突变是结核分支杆菌耐利福平的主要机理 ,PCR -SSCP可简便快速地检出rpoB基因的突变 ,有助于结核分支杆菌耐药性的快速检测和研究。

Objective:To construct a molecular drug sensitive method for detecting the drug tolerance of mycobacterium tuberculosis to RFP.Methods:The mutation of rpoB gene in mycobacterium tuberculosis by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP) was analysed,and compared with the results of drug-sensitive test.Results:Compared the PCR-SSCP picture to H37Rv,there had 81 mycobacterium tuberculosis resisted to RFP in 116,and 73's picture had difference in these 81 mycobacterium tuberculosis,but only 1 had difference in 35 mycobacterium tuberculosis that is senstive to RFP.The positive incidence is 90%,the specificity is 97.1%.Conclusion:The mutation of rpoB gene is the main mechanism that mycobacterium tuberculosis resisted to RFP,PCR-SSCP can detect the mutation of rpoB gene easily and quickly,it is helpful to detect and research the tolerance of the mycobacterium tuberculosis.