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Background. Congenital toxoplasmosis (CT) is an infectious disease caused by placental transfer of Toxoplasma gondii protozoa to the fetus, and can generate neurological and neurocognitive deficits, or death. As a mother’s infection poses the risk of fetal transmission in about 30% of the cases, appropriate preventive strategies are required for infection-related risk factors. The present study assessed the prevalence of T. gondii infection and the factors associated with congenital toxoplasmosis in pregnant women with assistance from the Public Health Service at Ouro Preto, Brazil.Methods. This cross-sectional study was conducted between April and December 2020. Pregnant women (n = 131) aged between 13 and 46 years, were recruited and evaluated for specific IgM and IgG antibody levels against T. gondii. These women also responded to a structured questionnaire, which was later used to determine the socioeconomic, environmental, gestational, clinical, and dietary patterns.Results. The prevalence of infection by T. gondii was 45.8% (n = 60); of these, multigravida women were more exposed to infection and were 2.6 times more likely to become infected with T. gondii compared to primigravida women (OR: 2.60; 95% CI: 1.25-5.39). A high prevalence of T. gondii seropositivity was identified in pregnant women in the region of Ouro Preto, Brazil, where part of the sample reported the absence of basic sanitation at home.Conclusion. Our data identified multigravida as a potential risk factor for gestational toxoplasmosis, based on the high positivity of anti-T. gondii serology in these women Thus, educational and preventive measures must be intensified in both primigravida and multigravida women, to increase awareness regarding the potential risks of contact with T. gondii.