This study surveyed the Toxoplasma (T.) gondii infection prevalence in the Korean rabbit population. Rabbits (n=142) were obtained from two breeding farms in the Gongju area, Chungnam Province, and in the Kochang area, Junbuk Province, Korea. Of 142 sera samples analyzed by enzyme-linked immunosorbent assay (ELISA), 15 (10.6%) exhibited T. gondii-specific IgG antibodies, and 1 (0.7%) rabbit harbored T. gondii-specific IgM. Female rabbits (9/84; 10.7%) had a similar T. gondii prevalence to males (6/58; 10.3%). When stratified by age, rabbits aged >1 year had a similar prevalence of T. gondii infection (7/66; 10.6%) to rabbits aged <1 year (8/76; 10.5%). Immunoblotting detected 6 major antigenic bands corresponding to T. gondii-positive sera at 20, 28, 30, 35, 63 and 77 kDa. Nested polymerase chain reaction (PCR) of whole-blood samples detected the T. gondii B1 gene in 23 rabbits (16.2%). All PCR-positive samples corresponded to partial T. gondii B1 gene sequences with 99% homology to a T. gondii sequence deposited in GenBank (accession number EU340874). Female rabbits (13/84; 15.5%) harbored a similar prevalence of T. gondii DNA to males (10/58; 17.2%). Rabbits aged >1 year had a similar prevalence (12/66; 18.2%) of T. gondii infection to rabbits aged <1 year (11/76; 14.5%). No statistically significant differences were observed regarding the prevalences of infection according to sex or age using molecular or serological tests. This study is the first survey using serological tests and nested PCR to analyze the T. gondii prevalence in rabbits in Korea.