Abstract: ObjectiveTo understand the hereditary character and evolutionary relationship of Brucella melitensis.MethodsThe multilocus sequence typing (MLST) assay was used to sequence 60 strains ofB. melitensis from Guangdong Province. Seven house keeping genes, 1 outer membrane gene, and 1 intergenic region of the 60 strains were sequenced and compared with alleles of standard strains to determine the sequence type. All the strains were analyzed through PCR detection, sequence and sequence analysis.ResultsThe sequences of all the 60 strains weregap（3）, aroA（2）, glk（3）, dnaK(2）, gyrB（1）, trpE（5）, cobQ（3）, omp25（8）, andInt-hyp（2）, consistent with the various alleles of sequences type ST8 reported in the literature. The similarity among the 60 local strains and 16 standard ones ranged from 2.5% to 100.0%. The strains with 100.0% similarity were divided into 12 ST types, of which the 60 local strains were ST8 type, same asB. melitensis 2 standard strain. ConclusionB. melitensis ST8 is dominant in Guangdong. MLST is only a supplementary method but cannot be used to differentiateB. melitensis in Guangdong Province.