【摘要】 目的 :调查深圳地区结核分枝杆菌耐利福平 (RFP)株rpoB基因突变的分布情况 ,建立结核分枝杆菌耐药基因快速检测的方法。方法 :对 5 5株结核分枝杆菌临床分离株的rpoB基因 2 80个碱基 (包括其核心区 75个碱基 )应用PCR 直接测序法 (PCR DS)测定序列 ,其中耐利福平株 5 1株 ,敏感株 4株。结果 :4株敏感株无突变 ,92 2 % (47/ 5 1)耐利福平临床分离株存在rpoB基因突变。基因突变导致 5 31位氨基酸突变率为 4 1 2 % (2 1/ 5 1) ;导致 5 2 6位氨基酸突变率为 2 9 4 % (15 / 5 1) ;导致 5 16位氨基酸突变率为 13 7%(7/ 5 1)。联合突变发生率为 2 0 % (1/ 5 1)。未检测到发生缺失或插入碱基突变的菌株。结论 :深圳地区结核分枝杆菌耐利福平株发生rpoB基因突变最常见的是 5 31位丝氨酸、5 2 6位组氨酸和 5 16位天冬氨酸的基因突变 ,三者突变率之和为 84 3% (43/ 5 1)。PCR DS方法可快速测定结核分枝杆菌RFP耐药基因突变。更多还原

【Abstract】 Objectives:To investigate the distribution of rpoB mutation in rifampin resistant clinical Mycobacterium tuberculosis isolates from Shenzhen,and to develop a fast method for detecting M.tuberculosis drug resistance.Methods:280 bp DNA fragment of rpoB genes,including 75 bp code region(rifampin resistant determination region),were sequenced by using PCR direct sequencing (PCR DS) techniques.There were 55 strains of M tuberculosis clinical isolates including 51 rifampin resistant strains and 4 rifampin sensitive strains.Results:92 2%(47/51) rifampin resistant trains had rpoB mutation in 75 bp code region.No mutation was found in rifampin sensitive strains.41 2(21/51) rifampin resistant strains had mutations located at 531 Ser;29 4%(15/51)strains had mutations at 526 His;13 7%(7/51)strains had mutations at 516 Asp。Combinative mutation rate was 2 0%(1/51)。There were not deletion or insertion mutation found.Conclusions:531 Ser ,526 His and 516 Asp were the main positions of mutation.The added mutation rate was 84 3%(43/51) PCR DS techniques should be a fast and reliable identified test for rifampin resistance in clinical isolates.更多还原