Abstract: Objective To understand the etiological characteristics of brucellosis and provide evidence for the diagnosis, treatment and control of brucellosis. Methods Blood samples collected from suspected brucellosis cases in Tongxiang to conduct rose-bengal plate agglutination test (RBPT) and standard tube agglutination test (SAT).The blood samples were taken from the positive persons to conduct bacterium enrichment by using biphasic blood culture bottles and further identification of the specie/type was conducted by using BCSP31-PCR and AMOS-PCR. Results One brucellosis case infected with Brucella melitensis biovar 3 was detected, the serum SAT result was 1:3200 (++). Epidemiological investigation and standard treatment was carried out for the patient. Conclusion The case was infected with Brucella melitensis biovar 3. It is suggested to conduct blood culture for the patient suffered from long time fever with unknown origin for the early detection and treatment of brucellosis.