[Objective] To establish the 16S rDNA sequence analysis method to identifiy Brucella and evaluate the specificity and feasibility of this method. [Methods] 16S rDNA were amplified from Brucella by polymerase chain reaction (PCR) method and the purified product were directly sequenced for further analysis. The 16S rDNA sequence of the bacteria that are known to cross-react serologically with Brucella were downloaded from the GenBank. DNAMAN was used for comparison of 16S rDNA sequence. [Results] Brucella 16S rDNA sequence similarity reached 99.74%, Brucella 16S rDNA sequence to serologically related bacteria had more pronounced differences. [Conclusion] 16S rDNA sequence analyses is a rapid, simple diagnostic and specific method.