结核分枝杆菌rpoB基因突变的研究

杨本付; 徐飚; 蒋伟利; 周佩源; 姜庆五

doi:10.11847/zgggws2005-21-08-13

结核分枝杆菌rpoB基因突变的研究

Study on rpoB gene mutations in Mycobacterium tuberculosis

摘要

摘要:
目的研究农村结核病人中利福平(RIF)耐药结核分枝杆菌(M.TB)临床分离株rpoB基因突变情况, 为农村结核病防制提供分子遗传学依据。
方法以标准菌株H37Rv为对照, 采用聚合酶链反应-单链构象多态性(PCR-SSCP)分析技术, 对来自苏北2个县2001年的145株M.TB分离株(其中敏感株117株, 耐RIF或含RIF耐多药株28株)的rpoB基因突变的发生率进行研究。
结果农村RIF耐药或含RIF耐多药M.TB临床分离株中rpoB基因突变的发生率平均为71.4%, 所有受试117株敏感株rpoB基因均无突变。与药敏试验结果对比PCR-SSCP试验的灵敏度为71.4%;其特异度为100%。
结论 M.TB耐RIF与rpoB基因突变密切相关, PCR-SSCP方法可检出RIF耐药M.TB的rpoB基因突变, 有助于M.TB耐药性的快速检测和耐药性防治。

Abstract:
Objective To understand rpoB gene mutaions in rifampin(RIF)-resistant Mycobacterium tuberculosis(M.TB)strains isolated from tuber culosis(TB)patients in rural area and to provide genetic evidence for TB control in rural areas.
Methods 145 isolates from two counties were examined for rpoB gene mutations by the polymerase chain reaction single strand conformation polymorphism(PCR-SSCP)method compared with virulent M.TB H₃₇R_v.
Results The prevalence of rpoB gene mutation was 71.4% in RIF-resistantor multidrug resistant M.TB from patients in this rural area.rpoB gene mutations were not found in all the 117 susceptible strains.In comparison with drug suscept ibility test results the sensitivity and specificity of PCR-SSCp assay were 71.4% and 100%, respectively.
Conclusion RIF-resistance of M.TB is strongly related to rpoB gene mutations.PCR-SSCP assay can detect rpoB gene mutations of RIF-resistant M.TB.It may thus be helpful to include a rapid detection of drug resistance of M.TB in resistance containment.

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