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

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。

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

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 non-tuberculous 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...