目的 建立布鲁氏菌基因缺失株荧光定量PCR方法，为其他布鲁氏菌毒株和基因缺失株鉴别诊断提供技术支持。 方法 根据GenBank中布鲁氏菌赤藓糖醇基因（eryA基因）设计合成特异性引物和探针，对引物、探针浓度、退火温度等反应条件进行优化后，检测该方法的特异性、敏感性和重复性。 结果 重组质粒标准品在1.56×10⁸ ~1.56×10 copies/μL呈良好线性关系；特异性试验结果显示，该方法对其他14种常见细菌病毒均无交叉反应；敏感性试验结果显示该方法对布鲁氏菌的检测敏感度为100 copies/μL。重复性试验结果显示，组内和组间变异系数均小于2%。 结论 TaqMan荧光定量PCR检测方法特异性强、灵敏度高、稳定性好，可用于布鲁氏菌基因缺失株和其他布鲁氏菌株的鉴别诊断。

Objectives A method for detecting the deletion strains of Brucella gene with TaqMan fluorescence quantitative PCR was established to provide technical support for the identification and diagnosis of other strains and deletion strains of Brucella gene. Methods Specific primers and probes were designed and synthesized based on the eryA gene of Brucella in GenBank. The specificity, sensitivity, and repeatability of the method established were tested after optimizing the reaction conditions including the concentration of primers and probes, and the temperature of annealing. Results The standard curve of recombinant plasmid had a good linear relationship between 1.56×10⁸ and 1.56×10¹ copies/μL. The results of specificity test showed that the method established had no cross-reaction with 14 other common bacterial viruses. The results of the sensitivity test showed that the method established had a detection sensitivity of 100 copies/μL for Brucella. The results of the repeatability test showed that the coefficient of variation within and between groups was less than 2%. Conclusions It is indicated that the method established has strong specificity, high sensitivity, and good stability, and can be used for the identification and diagnosis of the deletion strains and other strains of Brucella gene.