Very limited information is available concerning the epidemiology of T. gondii infection in pregnant women in eastern China. Therefore, a case-control study was conducted to estimate the seroprevalence of toxoplasmosis in this population group and to identify risk factors and possible routes of contamination. Serum samples were collected from 965 pregnant women and 965 age-matched nonpregnant control subjects in Qingdao and Weihai between October 2011 and July 2013. These were screened with enzyme linked immunoassays for the presence of anti-Toxoplasma IgG and anti-Toxoplasma IgM antibodies. 147 (15.2%) pregnant women and 167 (17.3%) control subjects were positive for anti-T. gondii IgG antibodies, while 28 (2.9%) pregnant women and 37 (3.8%) controls were positive for anti-T. gondii IgM antibodies (P = 0.256). There was no significant difference between pregnant women and nonpregnant controls with regard to the seroprevalence of either anti-T. gondii IgG or IgM antibodies. Multivariate analysis showed that T. gondii infection was associated with location, cats in home, contact with cats and dogs, and exposure to soil. The results indicated that the seroprevalence of T. gondii infection in pregnant women is high compared to most other regions of China and other East Asian countries with similar climatic conditions.