应用PCR方法扩增IS6110保守片段快速鉴定结核分支杆菌

Rapid identification of the Mycobacterium tuberculosis by polymerase chain reaction technique

目的 :了解PCR方法快速鉴定结核分支杆菌的可靠性 ,探索快速诊断结核病的实验方法。方法 :应用PCR方法扩增 149株结核分支杆菌临床分离株的IS6 110保守片段 ,并与结核分支杆菌H3 7Rv标准株进行比较。结果 :149株临床分离株中 ,140株可以扩增出与结核分支杆菌H3 7Rv标准株相同的 12 3bpDNA片段 (敏感性达 93 9%) ,而阴性对照均不能扩增出该片段。结论 :应用PCR方法扩增IS6 110保守片段来鉴定结核分支杆菌 ,结果准确可靠 ,且具有方便和快速等优点 ,有较大的临床应用价值。

Objective: To evaluate the reliability of rapid method to detect the DNA of Mycobacterium tuberculosis by polymerase chain reaction technique, and to develop a rapid technique to diagnose the infection of Mycobacterium tuberculosis. Methods: Mycobacterium tuberculosis strain H 37Rv was used as a control. A conserved IS6110 fragment of 149 Mycobacterium tuberculosis clinical isolates was amplified by polymerase chain reaction technique; and the PCR products was detected by agarose gel electrophoresis. Results: a clear 123 bp DNA band was detected in 140 of 149 isolates(93.3%) as well as the Mycobacterium tuberculosis strain H 37Rv, but no negative control displayed this band. Conclusion: The method of detecting the DNA of Mycobacterium tuberculosis by PCR amplifying the conserved IS6100 fragment is a reliable rapid way to diagnose the infection of mycobacterium tuberculosis.