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目的分析西安地区耐乙胺丁醇(EMB)结核杆菌临床分离株embB基因突变的特点,建立快速检测结核分枝杆菌乙胺丁醇耐药性的方法。方法应用聚合酶链反应-单链构象多态性(PCR-SSCP)和聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术检测104株结核分枝杆菌临床分离株的embB基因。结果以H37Rv标准株为对照,35株药物敏感株的SSCP和RFLP分析结果均与标准株一致,不存在突变;69株耐乙胺丁醇分离株中,39株(56.52%)SSCP泳动与标准株不一致,存在基因异常;19株(27.54%)RFLP分析存在基因异常。结论西安地区结核杆菌临床分离株对乙胺丁醇耐药与embB基因(尤其是306位密码子)突变有关,PCR-SSCP和PCR-RFLP技术可快速、准确地检测结核分枝杆菌对EMB的耐药性。
Objective To analyze the mutation of embB gene in clinical isolates of Mycobacterium tuberculosis (EMB) in Xi’an and to establish a rapid method for detecting the resistance of M. tuberculosis ethambutol. Methods The embB genes of 104 clinical isolates of Mycobacterium tuberculosis were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) . Results The results of SSCP and RFLP analysis of 35 drug-sensitive strains were consistent with those of the standard strains without H37Rv standard mutation. Of the 69 isoethachlor-resistant isolates, 39 (56.52%) SSCP motile and The results of RFLP analysis showed that there were gene abnormalities in 19 strains (27.54%). Conclusion The clinical isolates of Mycobacterium tuberculosis in Xi’an are related to the resistance to ethambutol to the mutation of embB gene (especially codon 306). PCR-SSCP and PCR-RFLP can detect Mycobacterium tuberculosis rapidly and accurately to EMB Resistance.