Objective To describe spatial distribution of brucellosis in Three-River Regions and research its epidemiological characteristics with the Geographic information technique. Methods We established Geographic Information System Database of brucellosis in Three-River Regions, epidemiological characteristics were overlay analyzed in infectious rate and prevalence rate with QHEndem ic-GIS and SPSS 17.0. Results Spatial characteristics of brucellosis were showed in GIS, the overlaid prevalence rates in Nangqian(2.00%/1.03%), Jiuzhi(0.81%/0.63%)and Dari(0.58%/0.33%)in inspection period were higher than that in inspection period. Overlaying the prevalence of brucellosis before and after inspection period, the prevalence show an upward trend. Conclusion GIS could reflect the state of brucellosis in Three-River Regions directly and correctly. Prevalence of brucellosis showed a sporadic phenomenon with spreading the neighborhood region, the prevalence showed an up-ward trend in these region, and provide scientific data of brucellosis prevention and control.