An antigen capture enzyme-linked immunosorbent assay (ELISA) was developed to detect bluetongue virus (BTV) in the vector Culicoides variipennis (Coquillett). This ELISA used polyclonal and monoclonal antibodies in a biotin-avidin immunoperoxidase system to detect BTV in individual adult female flies that had taken an infectious blood meal. Detection of BTV with the ELISA correlated with detection of BTV from the same flies by plaque assay in cell culture. The ELISA was group-specific for the five U.S. serotypes of BTV and did not cross react with closely related epizootic hemorrhagic disease virus.