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.