结核分枝杆菌rpoB、katG基因突变位点的克隆及其应用
发布时间：2018-08-14 15:23
【摘要】：目的：克隆结核分枝杆菌rpoB、katG常见基因突变位点,为核酸薄膜导流杂交技术快速检测临床标本结核分枝杆菌rpoB、katG基因突变提供阳性对照,以助判断临床标本结核分枝杆菌有无耐药。 方法：前期研究发现本地区结核分枝杆菌rpoB、katG基因常见的4个突变位点和6个突变形式,选择6株含有上述突变特征的临床分离株,PCR扩增其rpoB、katG基因片段进行T-A克隆,采用HindⅢ和EcorⅠ双酶切、PCR扩增及重组质粒测序鉴定重组质粒。核酸薄膜导流杂交技术检测此6个临床分离株的rpoB、katG基因片段。 结果：构建了6个含有rpoB、katG基因不同突变类型的重组质粒,三种方法鉴定明确重组质粒含有正确序列。核酸薄膜导流杂交技术检测6个临床分离株的rppB、katG基因片段结果显示在预期的位置显示阳性斑点。 结论：重组质粒可以作为核酸薄膜导流杂交技术检测临床标本时提示结核分枝杆菌有无耐药的稳定的阳性对照。
[Abstract]:Objective: to clone the common mutation sites of Mycobacterium tuberculosis rpoBhkatG gene, and to provide a positive control for rapid detection of rpoB katG gene mutation in clinical specimens by membrane diversion hybridization, in order to help determine the resistance of Mycobacterium tuberculosis in clinical specimens. Methods: four common mutation sites and six mutation forms of Mycobacterium tuberculosis rpoBnkatG gene were found in previous studies. Six clinical isolates with the above mutational characteristics were selected for T-A cloning by PCR amplification of the rpoBmkatG gene fragment of Mycobacterium tuberculosis. The recombinant plasmids were identified by Hind 鈪，
本文编号：2183292
[Abstract]:Objective: to clone the common mutation sites of Mycobacterium tuberculosis rpoBhkatG gene, and to provide a positive control for rapid detection of rpoB katG gene mutation in clinical specimens by membrane diversion hybridization, in order to help determine the resistance of Mycobacterium tuberculosis in clinical specimens. Methods: four common mutation sites and six mutation forms of Mycobacterium tuberculosis rpoBnkatG gene were found in previous studies. Six clinical isolates with the above mutational characteristics were selected for T-A cloning by PCR amplification of the rpoBmkatG gene fragment of Mycobacterium tuberculosis. The recombinant plasmids were identified by Hind 鈪，
本文编号：2183292
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