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目的探讨结核分枝杆菌异烟肼(INH)耐药的分子机制,建立快速检测结核分枝杆菌INH耐药性的方法。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测105株结核分枝杆菌临床分离株的katG基因,运用DNA测序和生物信息分析比对进行验证。结果以H37Rv标准株为对照,34株敏感株中5株(14.7%)存在KatG基因异常,105株耐药株中55株(52.4%)存在KatG基因异常。结论结核分枝杆菌对INH耐药与KatG基因突变有关,PCR-RFLP技术可快速、准确地检测结核分枝杆菌对INH的耐药性。
Objective To investigate the molecular mechanism of resistance to isoniazid (INH) in Mycobacterium tuberculosis and establish a rapid method for detecting the drug resistance of Mycobacterium tuberculosis INH. Methods The katG gene of 105 clinical isolates of Mycobacterium tuberculosis was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and verified by DNA sequencing and bioinformatics analysis. Results The H37Rv standard was used as a control. Among the 34 sensitive strains, KatG gene was abnormal in 5 (14.7%) and 55 (52.4%) in 105 resistant strains. Conclusion Mycobacterium tuberculosis resistance to INH is related to the mutation of KatG gene. PCR-RFLP can detect the drug resistance of Mycobacterium tuberculosis to INH rapidly and accurately.