PCR-DNA测序技术检测结核分支杆菌耐多药基因突变的研究  

Detection of the genomic mutations of the multidrug-resistant Mycobacterium tuberculosis by polymerase chain reaction-DNA sequencing

作　　者：熊国亮[1] 予季[1] 张慧慧[1] 刘珍琼[1] 

机构地区：[1]江西省胸科医院检验科,南昌330006

出　　处：《国际检验医学杂志》2008年第2期101-102,106,共3页International Journal of Laboratory Medicine

基　　金：江西省卫生厅立项资助项目(20045067)

摘　　要：目的应用PCR-DNA测序技术快速检测同时耐利福平和异烟肼结核分支杆菌分离株rpoB、KatG基因突变,评价其在检测结核分支杆菌耐多药性方面的价值。方法47株耐利福平和异烟肼结核分支杆菌临床分离株及30株结核分支杆菌敏感分离株用PCR-DNA测序技术分别检测rpoB、KatG基因突变。结果47株耐多药结核分支杆菌分离株中,检出43株rpoB基因突变,突变检出率为91.5%(43/47);31株KatG基因突变,突变检出率为66.0%(31/47);rpoB和KatG基因同时突变者31株,突变检出率为66.0%(31/47)。30株结核分支杆菌敏感株检出1株KatG基因突变。结论PCR-DNA测序技术方法敏感、准确、特异,可快速检测结核分支杆菌rpoB、KatG耐药基因突变,有利于耐多药结核分支杆菌耐药性的快速检测。Objective The aim of the study is to employe PCR-DNA sequencing to rapidly detect rpoB, KatG gene mutation, and to evaluate its clinical value in the detection of M. tuberculosis （MTB） multi-resistance. Methods The mutations of rpoB and KatG gene were detected with PCRDNA sequencing for 47 RFP- and INH-resistant MTB isolates and 30 RFP- and INH-sensitive ones. Results Among 47 multidrug-resistant MTB isolates, 43 isolates had rpoB gene mutation, and the mutation rate was 91.5% （43/47）; 31 isolates had KatG gene mutation, and the mutation rate was 66.0% （31/47）; 31 isolates had both rpoB and KatG gene mutations, and the mutation rate was 66.0% （31/47）. Of 30 drug sensitive MTB isolates, 1 strain had KatG gene mutation. Conclusion PCR-DNA sequencing is a seneitive, accurate and specific method for rapid detection of rpoB and KatG gene mutations of MTB, and useful for rapid detection of multidrug-resistant MTB.

关 键 词：分支杆菌 结核 基因 MDR 聚合酶链反应 突变 

分 类 号：R440[医药卫生—诊断学]