目的 :建立双重PCR技术快速鉴定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株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 2 4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。结果表明 ,用引物A1、A2扩增分支杆菌属共有的 6 5Kda抗原基因383bp片段和B1、B2扩增结核分支杆菌复合体特异的插入序列IS10 812 38bp片段鉴定结核与非结核分支杆菌是特异的。两对引物双重PCR检测结核分支杆菌DNA的敏感性分别为 1Pg和 10 0fg。双重PCR检测 34株临床分离株 ,结核分支杆菌可见 383bp和 2 38bp两条扩增带 ,非结核分支杆菌仅见 383bp一条扩增带。据此差别 ,能将结核与非结核分支杆菌鉴别开 ,并能在 1～ 2d完成试验。结论 :双重PCR技术为结核及非结核分支杆菌的快速鉴定提供了一个可供选择的手段 Objective: To establish dual PCR rapid identification of tuberculosis and non-tuberculosis mycobacteria. Methods: The optimal reaction conditions of double PCR were selected through experiments. The specificity of Mycobacterium bovis A1, A2 and B1, B2 was amplified and the sensitivity of Mycobacterium tuberculosis was amplified. The sensitivity and specificity of 34 Mycobacterium tuberculosis and Mycobacterium tuberculosis Clinical isolates were identified. Results: Primers A1 and A2 could amplify 24 mycobacteria tested, B1 and B2 only amplify Mycobacterium tuberculosis complex strains, and 12 pairs of primers amplified 12 kinds of non-mycobacteria were negative. The results showed that the identification of Mycobacterium tuberculosis and non-mycobacterium tuberculosis by using primers A1 and A2 to amplify the 383 bp fragment shared by Mycobacterium and the IS10 812 38 bp fragment specific to B1 and B2 amplification Mycobacterium tuberculosis complex . The sensitivity of the two pairs of primers for dual PCR detection of Mycobacterium tuberculosis DNA was 1 Pg and 10 0 fg, respectively. Twenty-four clinical isolates were detected by double-PCR. Two bands of 383 bp and 238 bp were found in Mycobacterium tuberculosis, and only one band of 383 bp was found in Mycobacterium tuberculosis. According to this difference, tuberculosis and non-tuberculous mycobacterium can be identified open, and can be 1 ~ 2d to complete the test. Conclusion: Dual PCR provides an alternative method for the rapid identification of tuberculosis and non-tuberculous mycobacteria