布鲁氏菌病与炭疽病多重PCR方法的建立

Establishment of Multiplex PCR Assay for Detection of Brucellosis and Anthrax

根据GenBank上已发表的布鲁氏杆菌BM21基因序列和炭疽杆菌PX01,设计合成两对特异性引物,分别建立了布鲁氏菌病(Brucellosis)和炭疽病(Anthrax)单项PCR诊断方法。通过对扩增条件的筛选,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和炭疽329bp特异性片段。通过对多重反应体系的特异性、敏感性实验和重复性试验,最终确定了布病和炭疽病的多重PCR诊断方法。

In this paper,two pairs of specific primers were designedand synthesized according to the gene sequence of Brucella BM21 and the plasmid PX01 of Bacillus anthracisthat were published in GenBank.Single PCR diagnostic methods were established for Brucellosis and Anthrax,respectively,and the optimum reaction conditions of the two primers were locked by the checkboard method.Simultaneously,specific PCR products of 255 bp for brucellosis and 329 bp for Anthrax were amplified.Through the multiple reaction system of specificity,sensitivity and repeatability experiments,the multiple PCR diagnostic methods were successfully established for brucellosis and anthrax.