TaqMan MGB双探针实时PCR快速检测结核分枝杆菌rpoB基因多位点突变方法的建立  

Rapid Detection of Rifampin Drug-resistant Gene Mutation of Mycobacterium tuberculosis by TaqMan MGB Dual-probe Real-time PCR

作　　者：王伟华[1] 刘伟[1] 

机构地区：[1]聊城市人民医院中心实验室,山东聊城252000

出　　处：《中华医院感染学杂志》2010年第3期324-326,共3页Chinese Journal of Nosocomiology

摘　　要：目的建立快速、灵敏的检测结核分枝杆菌rpoB基因常见突变位点的新方法。方法采用TaqMan小沟结合物(MGB)探针技术,检测146例活动性肺结核患者和35例非结核性肺部疾病患者痰标本中结核分枝杆菌rpoB基因突变情况,测序方法为参考方法。结果146例活动性肺结核患者痰标本中,双探针方法检出阳性118例,其中rpoB基因突变56例,与测序方法比较符合率100.0%;35例非结核性肺部疾病患者痰标本双探针方法检测均为阴性,特异性100.0%;该方法最低检出下限为1×103拷贝/ml。结论TaqMan MGB双探针方法能快速、准确地检测结核分枝杆菌rpoB基因位点突变情况,适用于临床对耐药结核分枝杆菌的快速诊断。OBJECTIVE To establish a new methed rapid, and sensitive detection of Mycobacterium tuberculosis rpoB gene mutation. METHODS M. tuberculosis rpoB gene mutation was detected by TaqMan minor groove binder （MGB） dual-probe real-time PCR with sputum of 146 active pulmonary tuberculosis patients and 35 nontuberculous pulmonary patients. Sequence analysis was regarded as a reference method. RESULTS Totally 146 sputum samples from active pulmonary tuberculosis patients were detected by TaqMan MGB dual probe method from them 118 were positive, of which rpoB gene mutation were found in 56 samples. It was competely accordant with the results by sequence analysis. The sensitivity of TaqMan MGB dual-probe was 1×10^3 copies/ml. CONCLUSIONS The method could detec the mutations of rpoB gene rapidly and accurately and could be used in diagnosis of rifampin drug-resistance in clinic.

关 键 词：结核分枝杆菌 利福平 RPOB基因 突变 

分 类 号：R378.911[医药卫生—病原生物学]