Bovine Leukemia Virus (BLV) is the agent of enzootic bovine leukosis (EBL), this disease is a neoplasm of lymphatic tissue in bovine species. The aim of this article was determinate the prevalence of bovine leukemia virus in dual-purpose cattle from nine municipalities that produce the 98% of milk in the Caquetá state, Colombia. Blood samples were collected in 100 herds dedicates to dual purpose cattle, obtained blood serum from 1000 animals, of which 893 corresponding to cows. Indirect Elisa test for detection of antibodies anti-GP51 of BLV was performance and the positive cases were considered if the serum-to-positive ratio with percentages of M/N lower than 40%. Categorized data were analyzed by contingency tables and ANOVA at the significant level of p<0.05 by DGC test was performed. The overall sero-prevalence of BLV in Caquetá state was of 25.18% (95%, CI: 21.9-28.46%), in males 26.25% and females 25.37% respectively. At municipal level the sero-prevalence varied of 7.12 to 41.81%. The prevalence of BLV at herd level was of 67% (95%, CI: 57.24-76.76%). In conclusion, the sero-prevalence of BLV in the dual-purpose livestock system over 36 months of age in Caquetá state is moderate, do not exist statistical difference between sero-prevalence of cows and bulls. At level of herds the prevalence of BLV is high. Improving strategies of control and managements in the herds, as well as implement policies of sanitary management are necessary.