Abstract: The seroprevalence of Toxoplasma gondii infection in pregnant women was investigated in Shandong Province, China between August 2010 and September 2013. A total of 7956 serum samples collected from 15 representative administrative regions in Shandong Province, China were evaluated by Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of specific antibodies. Overall, 13.33% (1218/9139; 95% CI: 12.63-14.02) of the samples was positive for specific IgG against T. gondii. The highest seroprevalence was found in low educational standards (15.77%; 533/3379; 95% CI: 14.55-17) and followed by secondary educational standards (12.15%; 454/3736; 95% CI: 11.1-13.2) and high educational standards (11.41%; 231/2024; 95% CI: 10.03-12.8). Furthermore, the seroprevalence of T. gondii was the highest in less more 8 weeks (time of pregnancy) (14.12%; 431/3053; 95% CI: 12.88-15.35), followed by 8-12 weeks (13.03%; 508/3900; 95% CI: 11.97-14.08) and >12 weeks (12.76%; 279/2186; 95% CI: 11.36-14.16). The present resulted indicated infection with T. gondii is prevalence in pregnant women in Shandong Province, China which provides relevant “base-line” data for conducting control strategies and measures against toxoplasmosis in this region and elsewhere in China. This is the first report of the comprehensive survey of T. gondii seroprevalence in pregnant women in Shandong Province, China.