布鲁氏菌病、结核病多重PCR方法的建立

Establishment of Multiplex PCR of Brucellosis and Bovine Tuberculosis

根据GenBank上已发表的布鲁氏杆菌BM21基因序列和结核分枝杆菌插入片段IS6110,设计合成两对特异性引物,初步建立了布鲁氏菌病(Brucellosis)和牛结核病(Bovine Tuberculosis)诊断方法,并以棋盘法锁定了两条引物的最适反应条件,同时扩增出布病255bp和牛结核129bp特异性片段。并通过对多重反应体系的特异性和敏感性实验,最终确定了布鲁氏菌病和牛结核病的复合PCR诊断方法。

Two pairs of specific primers were designed and synthesized according to the gene sequence of Brucella BM21 and the inserted segment IS6110 of Mycobacterium tuberculosis that were published in GenBank , the initial establishment of the method for diagnosis of brucellosis and bovine tuberculosis, and the optimum reaction conditions of the two primers were locked by the checkboard method. Simultaneously, specific PCR products of 255bp for brucellosis and 129bp for bovine tuberculosis were amplified. Through the multiple reaction system of specificity and sensitivity experiments, the composite PCR diagnostic method was successfully established for brucellosis and bovine tuberculosis.