耐多药结核分枝杆菌基因位点表达研究

Expression studies of gene loci for Mycobacterium tuberculosis

目的:了解临床分离的耐多药结核分枝杆菌菌株耐药基因的突变情况。方法:采用基因芯片法对耐多药结核分枝杆菌临床分离菌株的耐药基因进行检测,研究其耐药基因突变情况。结果:80株耐多药结核分枝杆菌临床分离株中未见单一耐药情况,全部耐异烟肼(isoniazid,INH)+利福平(rifampicin,RFP),其中耐异烟肼+利福平+链霉素(streptomycin,SM)+乙胺丁醇(ethambutol,EMB)的耐药率明显高于其他耐药情况,差异有统计学意义(P<0.05)。rpoB基因突变位点主要为第S531L、D516G、H526Y、D516V、H526D位密码子,其中第S531L和D516G位密码子突变率分别为72.5%和75.0%,明显高于其他密码子,差异有统计学意义(P<0.05)。katG基因突变位点主要为第315M位密码子,突变率为75%。inhA基因突变位点主要为第-15M位密码子,突变率为87.5%。rpsL基因突变位点主要为第43M、88M位密码子,突变率分别为85.2%、14.8%。embB基因突变位点主要为第306M2位密码子,突变率为65%。结论:PCR及基因位点突变测定可快速检测耐多药结核分枝杆菌的耐药基因,结核分枝杆菌耐多药性与多个耐药基因突变相关。

Objective To study the mutation of the resistance gene of the multi drug resistant Mycobacterium tuberculosis strains isolated from clinical. Methods The gene chip of drug resistance genes was used to examine the mutation of drug resistance gene of the clinical isolates of multi-drug resistant Mycobacterium tuberculosis. Results 80 strains of multi-drug resistant Mycobaeterium tuberculosis clinical isolates were not a single drug resistance situation, all strains were resistant to Isoniazid and Rifampicin at same time, the resistance rate of which resistant to Isoniazid＋Rifampicin ＋ Streptomycin ＋ Ethambutol was significantly higher than other kinds of drug resistance, the difference has statistical significance （P 〈 0.05）. rpoB gene mutations was mainly the S531L, D516G, H526Y, D516V,H526D codon, which the S531L and D516G codon mutation rates was 72.5%and 75%, significantly higher than the other codons, the difference have statistical significance （P 〈 0.05）. The mutation of katG gene was mainly 315M codon, the mutation rate was 75%. The mutation of the inhA gene was mainly-15M codon, the mutation rate was 87.5% .The mutation of rpsL gene was mainly 43M, 88M codon, the mutation rate was 85.2% and 14.8%. The mutation of cmbB gene was mainly 306M2 codon, the mutation rate was 65%. Conclusion The PCR and gene loci mutation analysis could quickly detect the gene of multi-drug resistant Mycobacterium tuberculosis, and the resistance to multi-drug resistant Mycobacterium tuberculosis was related to multiple mutations of resistance genes.