GRA2 is a highly immunogenic protein secreted from the dense granules of Toxoplasma gondii. Recent success in purifying full-length, soluble GRA2 from bacteria as a thioredoxin (TRX)-(Hisx6) fusion protein led to investigate the antigenicity of the recombinant protein against human sera. On immunoblots, TRX-(Hisx6)-GRA2 was recognized by sera collected in Iran from T. gondii-infected pregnant women. An IgG enzyme-linked immunosorbent assay was developed to evaluate the reactivity of sera, collected from pregnant women both in France and Iran, to the TRX-(Hisx6)-GRA2 fusion protein. Specificity of the test was 96.4%. Sensitivity of the GRA2 enzyme-linked immunosorbent assay ranged from 95.8% (sera collected in France) to 100% (sera collected in Iran) for sera of acute infection and from 65.7% (sera collected in France) to 71.4% (sera collected in Iran) for sera of chronic infection. The recombinant GRA2 could thus advantageously complement previously described T. gondii antigens for the serodiagnosis of acute Toxoplasma infection.