To investigate the seroprevalence of Toxoplasma gondii infection in domestic cats in central China, 843 serum samples were collected in Henan province between March 2015 and May 2016 and tested for IgG antibodies against T. gondii using the enzyme-linked immunosorbent assay (ELISA). The overall seroprevalence of T. gondii was 21% (178/843). No significant difference was observed based on the sex of cats (p > 0.05). Significantly higher seroprevalence (p < 0.05) was observed in mixed-breed cats (24%) compared to purebred cats (17%). Seroprevalence in rural cats (29%) was significantly higher (p < 0.01) than in urban cats (16%), and increased significantly (p < 0.01) with age. These results showed that T. gondii was highly prevalent in domestic cats in Henan province, central China, which might have important implications for public health.