In this study, a dot-immunobinding assay (DIA) using nitrocellulose membrane was performed for the detection of Brucella antibodies in 206 sheep sera. Extract of Brucella abortus strain S99 obtained by heat treatment was used as antigen. The results were compared with those of Rose Bengal plate agglutination test (RBPAT). Of the 96 sera collected from the 8 aborted sheep flocks, 46 were positive by both tests and 17 were positive only in DIA. Of the 110 sera collected from slaughtered sheep, 13 were positive by both tests and 4 were positive only in DIA. Of the total 206 Sera tested, 147 were negative by RBPAT and only 126 were negative in DIA. All the sera giving positive reactions in RBPAT were also positive in DIA. it is concluded that DIA would be useful to apply for the detection of Brucella antibodies in sheep sera.