Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis. This study was conducted to clarify the seroprevalence of BLV in the Republic of Korea. Blood samples were obtained from Korean native cattle farms in all provinces of South Korea except Jeju. A commercial indirect enzyme-linked immunosorbent assay with 4,498 samples to detect antibodies against BLV was conducted. The results revealed that the prevalence of BLV was dependent upon age, with increasing prevalence among cattle occurring until they were 5 years old. The highest seroprevalence in cattle was observed in Chungnam (29.6%) and the lowest was observed in Jeonnam (2.6%). The mean overall prevalence for BLV antibodies in the survey was 10.2%, indicating that BLV is widespread nationwide.