A study was conducted to investigate the serological prevalence of Border Disease Virus (BDV) antibodies in newly introduced Dorper sheep flocks at Debre Birhan Agricultural Research Center (DBARC), Amhara Region, Ethiopia. Blood samples were collected from the jugular vein of 186 sheep in the research center and the serum was decanted and sent to the National Animal Health Diagnostic and Investigation Center, Sebeta, Ethiopia for laboratory examination. All serum samples were examined using blocking ELISA test with a sensitivity of 94.3% and specificity of 93.7% to detect antibodies for BDV. Of these, 55 (29.57%) tested sero-positive and the rest 131 (70.43%) tested sero-negative for BDV antibodies. The sero-prevalence of sheep born in the research center was found at 23.97% (29/121) whereas the sero-prevalence of those sheep imported directly from abroad was found at 40% (26/65). In this research, a higher prevalence of BDV antibodies were recorded in the imported sheep than in the sheep which are born in the research center and the difference in prevalence between the two groups was found statistically significant (P<0.05). Therefore, it is of utmost importance to study the status of the disease from indigenous breeds of sheep and utilize more sensitive tests such as RT-PCR.