[Background] Brucella canis is a pathogenic bacterium of canine brucellosis and mainly leads dogs to abortus and reproductive disorders. B. canis is a rare source of infection for human brucellosis, but safety risk of human posed by B. canis remains controversial. At present, the epidemiology characteristics and genetic diversity of B. canis research in China were lacking relatively. It is significant to investigate epidemiology characteristics and genetic diversity of canine brucellosis for strengthening surveillance and control of canine brucellosis. [Objective] Epidemiology characteristics and genetic diversity of B. canis strains were investigated in order to provide a reference for control and prevention of canine brucellosis. [Methods] Both conventional bio-typing methods and BCSS-PCR were applied for species identified of 63 B. canis strains. The genetic diversity of B. canis strains were investigated by the means of HGDI. MLVA method was used for cluster analysis of B. canis to reveal the epidemiological characteristics of canine with BioNumerics 5.0 software. goeBURST software was applied to construct minimum spanning tree of B. canis strains for elucidate characteristics of geographic origin of B. canis strains in China. [Results] Both conventional bio-typing methods and BCSS-PCR were used and 63 tested strains were all B. canis, and 100% identification coincidence rate was found between conventional bio-typing method and BCSS?PCR assay. The analytical sensitivity of BCSS-PCR at DNA sample was 10?3 (50 pg/μL DNA of detecting B. canis). The higher genetic diversity was observed in B. canis strains in China. The results of genetic diversity investigated have showed that five loci in Panel 2B presented higher variability, the orders of allele genotypes from high to low were bruce09 (11), bruce07 (8), bruce16 (7), bruce04 (6), bruce30 (5). MLVA cluster analysis showed that there were three small-scale outbreaks of canine brucellosis in Beijing and sporadic in other areas. B. canis strains in this study were divided into five geographic groups (I?V), MLVA-11 genotype 26 clone group was predominated population. The strains of this clone group had common geographic origin with the strains from countries such as the United States, Greece, Canada, France, Romania and Korea. The others four geographic groups were unique lineages of B. canis in China. [Conclusion] There were higher genetic diversity and extensive of geographic origin of B. canis strains observed in China, and the results exhibited characteristics of origin and evolution of co-existing of imported and native specific lineage.