Rose bengal plate test (RBPT) antigens from Brucella melitensis and Brucella suis S 2 were prepared and compared with RBPT antigen prepared from classical Brucella abortus S 99. A total of 54 sera samples, of which 7 were collected from humans with brucellosis and 47 from infected sheep, were studied by using RBPT and serum agglutination test (SAT). RBPT and SAT results showed that RBPT antigens prepared from Br. melitensis and Br. suis S 2 were compatible with RBPT antigen prepared from Br. abortus S 99. Therefore, we concluded that RBPT antigens can be prepared from Br. melitensis and Br. suis S 2 and used for epidemiological surveilance of human and sheep brucellosis.