合肥地区耐多药结核分枝杆菌异烟肼、链霉素及乙胺丁醇耐药相关基因位点表达研究

Expression studies of drug-resistence related genes loci of isoniazid,streptomycin and ethambutol in multi-drug resistant Mycobacterium tuberculosis in Hefei area

目的 研究耐多药结核分枝杆菌异烟肼(INH)、链霉素(SM)及乙胺丁醇(EMB)耐药相关基因位点的表达情况。方法 选取该院2013年1月至2017年12月临床分离的101株耐多药结核分枝杆菌,利用测序法和基因芯片法分别检测INH、SM和EMB耐药相关基因及突变位点。结果 101株耐多药结核分枝杆菌中对SM耐药率69.3%;对EMB耐药率51.4%;对SM和EMB二者均耐药的耐药率44.5%。基因芯片法检出95例INH耐药相关基因突变,其中katG基因315M位点突变率为77.8%;inhA基因-15M位点突变率为89.4%。检出64例rpsL SM耐药相关基因,其中43M位点突变率为89.0%;88M位点突变率为11.0%。检出47例embB EMB耐药相关基因,以306M2位点突变为主,突变率为68.0%。测序法检出98例INH耐药相关基因突变,katG基因突变以315位点为主突变率为73.4%;inhA基因突变以-15位点为主突变率为 87.7 %;oxyR-ahpC基因-12和-10位点突变率分别为10.2%和8.2%。检出68例SM耐药相关基因突变,rpsL基因以43位点为主突变率为86.8%;rrs基因512、513、516位点突变率分别为5.9%、8.8%、8.8%。检出50例EMB耐药相关基因embB以306位点为主突变率为88.0%。结论 耐多药结核分枝杆菌INH、SM及EMB耐药基因位点表达可作为结核分枝杆菌临床检测耐药性的有效依据,为提高耐多药结核分枝杆菌耐药性检测的敏感度,应将INH耐药相关oxyR-ahpC基因和SM耐药相关rrs基因纳入我国快速分子诊断产品中。

Objective To study the expression situation of drug-resistence related genes of isoniazid,streptomycin and ethambutol in multi-drug resistant Mycobacterium tuberculosis. Methods 101 strains of clinical isolated multi-drug resistant Mycobacterium tuberculosis selected in a hospital from January 2013 to December 2017 were separately analyzed by sequencing method and gene chip to detect the isoniazid,streptomycin and ethambutol resistance related genes and mutations loci. Results The resistance rate against streptomycin in 101 isolated multi-drug resistant Mycobacterium tuberculosis stains was 69.3%;Resistance rate aginst ethambutol was 51.4%,resistance rate against streptomycin and ethambutol was 44.5%;95 cases of gene mutations related to isoniazid were detected by gene chip method,among which the mutation rate of katG gene at 315M was 77.8%,the mutation rate of inhA gene at -15M was 89.4%;The mutation rate at 43M loci was 89.0 % in 64 cases of streptomycin resistance related gene rpsL.The mutation rate at 88M site was 11.0%;47 cases of embB were detected to be drug resistance gene related to ethambutol,mainly with 306M2 site mutation rate of 68.0%.98 cases of gene mutation of isoniazid resistance were detected by sequencing method,and the mutation rate of katG was 73.4%,mainly at 315 loci;The mutation rete of inhA gene was 87.7% at -15 loci;the mutation rate of oxyR-ahpC gene were 10.2% and 8.2% at oxyR-ahpC -12 and -10 site.68 cases of streptomycin resistance gene mutation were detected,and the mutation rate of rpsL gene mainly at 43 loci was 86.8 %,the mutation rates of RRS gene mainly at 512,513,516 site were 5.9%,8.8% and 8.8%.The major mutation rate of embB at 306 loci related to ethambutol resistance in 50 cases was 88.0%. Conclusion The expression of resistance gene loci of isoniazid,streptomycin and ethambutol can be regarded as an effective basis of clinical detection of drug resistance in multi-drug resistant Mycobacterium tuberculosis.Isoniazid related resistant oxyR-anpC gene and streptomycin related resistant RRS gene should be included in China′s rapid molecular diagnosis products in order to improve the sensitivity of multi-drug resistant Mycobacterium tuberculosis drug resistance detection.