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

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.