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为建立一种比现有方法敏感、准确性高、重复性好的结核分枝杆菌DNA定性定量检测方法 ,以TaqMan探针技术为基础 ,运用TaqMan MGB探针 ,实时检测临床标本中的结核分枝杆菌DNA .用来自临床标本的DNA及克隆于载体的IS6 1 1 0序列检测所建立方法的有效性 .结果显示 ,所建立方法的最低检测限度为 1个基因拷贝 反应 ,在每反应 1 0 0 ～ 1 0 8拷贝范围内 ,Ct 值同DNA量的对数呈线性关系 .同一模板不同时间或同一时间不同管内扩增 ,所得Ct 值恒定 .用该方法检测 37例结核分枝杆菌培养阳性的痰液标本 ,敏感度为 1 0 0 % ;用该方法检测 1 6例TB系列阴性参考品 ,特异性为1 0 0 % .结果表明 ,所建立的方法是用于结核分枝杆菌定性定量检测较理想的方法
To establish a qualitative and quantitative detection method of Mycobacterium tuberculosis that is more sensitive, accurate and reproducible than the existing methods, based on the TaqMan probe technology, TaqMan MGB probe is used to detect tuberculosis in clinical samples in real time The results showed that the minimum detection limit of the established method was 1 gene copy reaction, and the reaction time was 1 0 In the range of 0 ~ 108 copies, the Ct value was linear with the logarithm of the amount of DNA.The same template was amplified in different tubes at different times or at the same time, and the Ct value was constant.This method was used to detect 37 cases of Mycobacterium tuberculosis culture positive Of sputum specimens with a sensitivity of 100%. The detection of 16 TB-negative samples by this method was 100% specific. The results showed that the method was established for the qualitative and quantitative identification of Mycobacterium tuberculosis Detection of the ideal method