The prevalence of Chagas disease has decreased in the Americas region due to vector control measures. However, non-vectorial transmission through blood transfusions and organ transplantation has gained importance in recent years. Screening among blood and organ donors are essential to reduce Trypanosoma cruzi transmission and could provide information to estimate population prevalence. We conducted a cross-sectional study on the prevalence of immunoglobulin G (IgG) antibodies against T. cruzi in healthy blood donors, solid organ donors and heart transplant recipients from 2012 to 2019. We found a total of 99 357 IgG T. cruzi results during the study period. The cumulative seroprevalence in healthy blood donors was 0.13% (95% confidence interval (CI) 0.10–0.15), in organ donors was 0.53% (95% CI 0.06–1.92) and in heart transplant recipients was 3.03 (95% CI 0.07–15.75). Seroprevalence trend in healthy blood donors showed annual increase between 2012 and 2015, decreasing in the following years. No trend was seen in organ donors neither heart recipients. Adjusted rates did not show difference by sex and age among blood donors. No significant increases in seroprevalence T. cruzi were found during the study period. T. cruzi transmission remains low.