某地区结核分枝杆菌利福平和异烟肼耐药相关基因突变特征分析  

The characterization of gene mutation in rifampin and isoniazid resistant mycobacterium tuberculosis isolated from a city

作　　者：杨辉[1,2] 张国良[2] 张明霞[2] 陈心春[2] 陈建波[2] 吴爱武[3] 

机构地区：[1]广州医学院医学检验系,510182 [2]广东省深圳市第三人民医院肝病研究所,518112 [3]广州医学院第一附属医院检验科,510120

出　　处：《重庆医学》2012年第34期3591-3593,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(81172732)

摘　　要：目的了解深圳地区结核分枝杆菌临床分离株利福平和异烟肼的耐药相关基因的突变特点。方法 60株结核分枝杆菌临床分离株通过全自动快速分枝杆菌培养鉴定系统(BACTEC-MGIT960)进行鉴定和药敏分析。在这60株菌株中,针对包括利福平耐药决定区(RRDR)在内的rpoB基因和异烟肼耐药相关位点katG315、inhA-15、kasA269、kasA312进行PCR扩增,并将扩增产物T-A克隆后进行测序。结果在60株结核分枝杆菌临床分离株中,检出利福平耐药株45株,敏感株15株;异烟肼耐药株41株,敏感株19株;利福平和异烟肼同时耐药的有14株。在45株利福平耐药株中,rpoB基因RRDR突变发生率为91.1%(41/45),共检测到12种突变形式,主要突变位点在531、526、516、533,以Ser531Leu突变最常见,占rpoB发生基因突变株的51.2%(21/45)。在41株异烟肼耐药株中,katG315位点有29株发生突变,包括28株katG315(AGC-ACC)和1株katG315(AGC-AAC)突变,突变发生率为70.7%(29/41);inhA-15位点有9株发生突变,均为(C-T)突变,突变发生率为21.95%(9/41)。kasA基因未发现常见突变形式。结论深圳地区结核分枝杆菌利福平和异烟肼耐药相关基因突变位点与国内外研究基本一致,但各位点频率有所不同。Objective To investigate the characterization of gene mutation in Rifampin and Isoniazid resistant Mycobacterium tu- berculosis isolated from Shenzhen. Methods 60 Mycobacterium tuberculosis clinical strains were identified and their drug resistance in phenotype were tested by BACTEC MGIT960, rpoB, katG315, inhA 15, kasA269 and kasA312 genes were amplified by PCR and the fragments were cloned into T vector and sequenced. Results Among the 60 Mycobacteria tuberculosis clinical strains,45 rifampicin resistant strains and 15 rifampicin susceptible strains;41 isoniazid resistant strains and 19 isoniazid susceptible strains;14 multiple drug resistance strains were detected. About 91.1% （41/45） of Mycobacterium tuberculosis isolates in 45 rifampicin drug resistant strains has mutations in rpoB and there were 12 different mutations,gene locus 531,526,516,533 mutation had a higher frequency especially gene locus 531 which had 51.2% （21/41） of mutational rate in 41 strains with rpoB gnen mutation. Point mutations at gene locus katG315 including katG315 （AGC-ACC）in 28 isoniazid resistance strains and katG315 （AGC AAC） in 1 isoniazid resistance strain, were found in 70.7% （29/41） of isoniazid resistant strains. 21.95% （9/41） of Mycobacterium tuberculosis isoniazid resistant strains had inhA-15 （C-T）mutations, no mutation in kasA was found in the study. Conclusion Ompared with other studies,the mutational gene site of Mycobacterium tuberculosis isolates with drug esistance of rifampicin and isoniazid in Shenzhen were consistant,but the gene mutation frequency in common gene locus of drug esistance were different.

关 键 词：分枝杆菌 结核 药物耐受性 突变 

分 类 号：R440[医药卫生—诊断学]