Four white-tailed deer (Odocoileus virginianus) were experimentally inoculated with Borrelia burgdorferi to determine serologic response by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Deer had antibodies by ELISA by 2 to 3 wk post-inoculation (PI) and remained positive for 10 wk. Deer demonstrated immunoblotting reactivity between 10 and 14 days PI and consistently showed antibody response to nine B. burgdorferi antigens. Attempts were made to recover the spirochete from blood and tissues; B. burgdorferi was isolated from an ear punch biopsy from one of the inoculated deer.