Background: Toxoplasmosis during pregnancy can causes adverse outcomes. Regarding its importance in prenatal care, seroprevalence of Toxoplasma gondii infection among pregnant women was studied in Ardabil City, during 2021-2022. Methods: In a cross–sectional study with a cluster sampling, 244 pregnant women that attended in health care centers of Ardabil City, Ardabil Province, Iran in 2021-2022 were selected. Serum samples were collected and Anti- Toxoplasma IgM (immunoglobulin M) and IgG (immunoglobulin G) was detected. Furthermore, a questionnaire filled for all participants during samples collection and risk factors assessed. Data were analyzed by SPSS software. Results: Participant's age ranged from 16 to 43 years with average of 23± 5.24 years. IgG anti- T. gondii antibody was detected in 22.1% of pregnant women (54/244). In none of participants IgM anti- Toxoplasmosis antibody was detected. There was no significant relationship between demographic variables and risk factors of toxoplasmosis with serology test results. Conclusion: Approximately, 77.9% of pregnant women had no antibody against the T. gondii infection. Therefore, health education, counseling of pregnant women and screening in high-risk pregnant women is recommended to prevent from fetal complications.