应用双重PCR技术快速鉴定结核与非结核分支杆菌

目的：建立双重ＰＣＲ技术快速鉴定的结核与非结核分支杆菌。方法：通过试验选择双重ＰＣＲ最佳反应条件，检测引物Ａ１，Ａ２和Ｂ１，Ｂ２扩增分支杆菌的特异性及扩增结核分支杆菌的敏感性，并对３４例结核分支杆菌和非结核分支杆菌临床分离株鉴定。结果：引物Ａ１，Ａ２能扩增所试２４种分支杆菌，Ｂ１，Ｂ２仅扩增结核分支杆菌复合体菌种，两对引物扩增１２种非分支杆菌均为阴性。

Application of duplexing PCR in rapid detecuon for tuberculous and nontuberculous mycobacteria

Objective:To establish a duplexing PCR technique to detect tuberculous and nontuberculous mycobacteria rapidly,Methods:The best reaction parameters of duplexing PCR,the specificity and sensitivity of the PCR primers A1,A2,and B1,B2 were checked by preexperiments. And then 34 cell strains of tuberculous mycobacteria and nontuberculous mycobacteria were identified.Results:Primer A1 and A2 could amplify al the 24 kinds of the experimental mycobacteria,while primer B1 and B2 could only amplify the complexus strain of tuberculous mycobacteria.The results showed that the sensitivities of this deuplexing PCR using two pairs of primers were 1pg and 100fg respectively,The duplexing PCR showed tow electrophoretic bands of 383bp and 238bp for tuberculous mycobacteria, but just one electrophoretic band of 383bp for nontuberculous mycobacteria, This result could differentiate tuberculous mycobacteria from nontuberculous mycobacteria within 1-2 day. Conclusions:The duplexing PCR technique provides a valuable method for rapidly identifyling tuberculous mycobacteria and nontuberculous mycobacterial.