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Background: Seroprevalence and incidence of toxoplasmosis in women of child bearing age has remained a contentious issue in the Indian subcontinent. Different laboratories have used different patient recruitment criteria, methods and variable results, making these data difficult to compare. Aim: To map the point-prevalence and incidence of toxoplasmosis in India. Material and Methods: In this cross-sectional study, a total of 1464 women of fertile age were recruited from 4 regions using similar recruitment plans. This included women from northern (203), southern (512), eastern (250) and western (501) regions of India. All samples were transported to a central laboratory in Delhi and tested using VIDAS technology. Their age, parity, eating habits and other demographic and clinical details were noted. Results: Most women were in the 18–25 years age group (48.3%), followed by 26–30 years (28.2%) and 31–35 years (13.66). Few (45) women older than 35 yr. were included. Overall prevalence of anti-Toxoplasma IgG antibodies was seen in 22.40%, with significantly more in married women (25.8%) as compared to single women (4.3%). Prevalence increased steadily with age: 18.1% in the 18–25 yr. age group to 40.5% in women older than 40 yr. The prevalence was high (66%) in those who resided in mud houses. Region-wise, the highest prevalence was observed in South India (37.3%) and the lowest (8.8%) in West Indian women. This difference was highly significant (P<0.001). Prevalence was 21.2% in East India and 19.7% in North India. The IgM positivity rate ranged from 0.4% to 2.9% in four study centers. Conclusions: This pan-India study shows a prevalence rate of 22.4% with a wide variation in four geographical regions ranging from as low as 8.8% to as high as 37.3%. The overall IgM positivity rate was 1.43%, indicating that an estimated 56,737–176,882 children per year are born in India with a possible risk of congenital toxoplasmosis. Author Summary: Toxoplasmosis is a protozoan parasitic disease commonly transmitted and propagated by cats as family pets. Infection acquired during pregnancy can lead to congenital abnormalities in the fetus, still birth or intrauterine death. Seroprevalence and incidence of toxoplasmosis in Indian women of child bearing age has remained a contentious issue. Different laboratories have used different patient recruitment criteria, methods and variable results, making these data unreliable. There is no published pan-India seroprevalence study. Hence, a seroprevalence study was undertaken comprising 1464 women of reproductive age representing four distinct geographical regions of India. This resulted in an estimated prevalence of 22.4% (328); the highest prevalence being in South India (37.3%) followed by East India (21.2%) and North India (19.7%). West Indian women had the lowest seroprevalence (8.8%). This difference was highly significant. In our analysis we determined the possible risk-factors of infection in these women. These included lower socioeconomic status, residing in mud plastered houses, consumption of raw salad, drinking untreated water, owning pets and advanced age. Overall, the incidence rate of toxoplasmosis was 1.43%. Extrapolating the data, we estimate that between 56,737 and 176,882 children a year may be born in India with a possible risk of congenital toxoplasmosis, which can manifest itself in-utero or several years after birth. [ABSTRACT FROM AUTHOR]