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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。
Objective: To investigate the distribution of rpoB gene mutation in Mycobacterium tuberculosis (RFP) strain of Mycobacterium tuberculosis in Shenzhen and to establish a rapid detection method of Mycobacterium tuberculosis resistance gene. Methods: Two hundred and fifty base pairs of rpoB gene (including 75 nucleobases in core region) of 55 strains of Mycobacterium tuberculosis clinical isolates were sequenced by PCR direct sequencing (PCR DS), of which rifampicin 5 1 strain, 4 sensitive strains. Results: No mutation was found in the four strains. The rpoB gene mutation was found in 92 2% (47/51) rifampin clinical isolates. The mutation rate of 5 31 amino acids was 4 1 2% (2 1/5 1); the mutation rate of 5 2 6 was 2 9 4% (15/5 1); the mutation rate of 5 16 amino acids was 13 7% (7/5 1). The incidence of co-mutation was 20% (1/5 1). No deletions or insertions of base-mutated strains were detected. Conclusion: The most common mutations of rpoB gene in Mycobacterium tuberculosis rifampin resistant strains in Shenzhen are the mutation of 5 31 serine, 5 2 6 histidine and 5 16 aspartic acid, the mutation rate of the three And 84 3% (43/5 1). PCR DS method for rapid determination of M. tuberculosis RFP resistance gene mutations.