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Congenital toxoplasmosis can cause severe consequences in the fetus, such as spontaneous abortion which is affected by parasite strain. Also, recent studies revealed the high genetic diversity of Toxoplasma gondii. This study aims to investigate the serological status of T. gondii in pregnant women, multilocus genotyping in aborted fetuses’ tissue, and archived formalin‐fixed paraffin-embedded placenta. This study was performed on 100 pregnant women with spontaneous abortion and their aborted fetuses, and 250 of the archived placentae in Iran. The blood and tissue were examined for seroprevalence and genotype determination of T. gondii using ELISA and multilocus nested-PCR–RFLP, respectively. Anti-T. gondii IgG and IgM were detected in 68 samples (68%) and 1 (1%) out of 100 serums. Toxoplasma DNA was identified in 1 (1%) aborted fetuses’ tissue and 32 (12.8%) placenta samples. Overall, ten …