快速检测手部非结核分枝杆菌感染的多重聚合酶链反应的方法建立

Establishment of multiplex PCR method for rapid detection of nontuberculous mycobacteriums infection in the hand

目的建立一种可快速检测手部常见非结核分枝杆菌感染的具有特异、敏感度高的多重聚合酶链反应（PCR）方法。方法应用引物设计软件以海、鸟、堪萨斯、偶发分枝杆菌的特异靶序列构建多重PCR方法，用上述非结核分枝杆菌标准菌株的DNA检测每一细菌的单一PCR和构建的多重PCR特异性，并测序对比评估验证多重PCR引物的特异性。用该方法鉴定26份临床标本。结果检测26例临床标本，阳性检出达7／8以上，鉴定时间较传统方法短。结论本研究方法可快速、特异、敏感有效的检测出常见手部非结核分枝杆菌感染，可用于临床非结核分枝杆菌感染的鉴定。

Objective To establish a multiplex polymerase chain reaction （mPCR） method with high sensitivity and specificity for rapid detection of common nontuberculous mycobaeterium （NTM） infection in the hand. Methods Application of primer design software to the mycobacterium marinum, mycobacterium avium, mycobacterium kansasii and mycobacterium fortuitum, the specific gene sequences were used to design construction of multiplex PCR and detection of DNA from the non tuberculous myeobacterial standard strains of each bacterium of single PCR and multiplex DNA accuracy and sequence contrast evaluation to verify the specificity of multiple PCR primers. 26 clinical specimens were identified by this method. Results Detection of 26 cases of clinical samples, positive detection of more than 7/8, the identification time is shorter than the traditional method. Conclusion The research method can be rapid, specific, sensitive and effective to detect the common hand of mycobacterium tuberculosis infection, can be used for clinical identification of mycobacterium tuberculosis infection.