The objective of this study was to estimate the seroprevalence of anti–Toxoplasma gondii antibodies and DNA of women with spontaneous abortions in 2 hospitals located in Yucatan, Mexico. Between June 2008 and May 2009, blood samples were taken from 100 women with spontaneous abortion attending the Ticul City Communitarian Hospital and the Merida Mother–Kid Hospital. The sera were tested for anti–T. gondii IgG and IgM antibodies. Blood samples (5 ml with anticlotting agent) were also used for polymerase chain reaction (PCR) assay, to detect T. gondii DNA. Forty-two of the 100 samples were negative. Of the positive samples (n = 58), 32 were positive to IgG, 2 to IgM, 5 to IgG and IgM, 6 to IgG and PCR, 1 to IgM and PCR, and 12 to IgG, IgM, and PCR. Accordingly, 55% of the women were seropositive to at least IgG, 20% to at least IgM, and 19% via PCR. Differences between hospitals were significant (P < 0.05) only for IgM. The risk of infection (IgM positive) was 2.85 (odds ratio [OR] 95%, confidence interval [CI]; 1.03–7.87) times greater in women patients at the Merida Mother–Kid Hospital, than those at the Ticul Communitarian Hospital. More studies are needed to evaluate the impact of this disease and to establish strategies to follow in order to reduce congenital toxoplasmosis in the populations at risk.