Introduction: Transfusion is a mechanism of transmission of Chagas' disease. There are no studies of the costs of the screening test in Colombian blood-banks. Objective: To estimate the costs related to a screening test for Chagas in blood-donors in two Colombian blood-banks, 2015. Materials and methods: A micro-costing study from the perspective of the health care provider was conducted to estimate the cost of Chagas test in two blood-banks, Banco de Sangre Cruz Roja Seccional-Bolivar and Banco de Sangre Hospital de Yopal-Casanare , for four cost categories: 1. Administrative costs: public services and insurance costs were calculated based on the blood-bank area in square meters; 2. Capital costs: building and equipment costs that were annualized using a 3% discount rate and a lifespan of 20 years for building and 5 for equipment. 3. Costs of Chagas test materials and reagents, adjusted by blood-bank production level and 4. Costs of staff who participated in Chagas test processing. Costs were reported with and without the cost of transfusion bags and immunohematology tests. Results: The cost of Chagas test in Seccional-Bolivar was COP$ 37,804 (US$12), and the blood bag and immunohematology test cost COP$25,941 (US$8.2) and COP$6,800 (US $2.2), respectively. In Yopal-Casanare, the costs were COP$77,384 (US$24.6), COP$30,141 (US$9.6) and COP$12,627 (US$4), respectively. Personnel cost accounted for the highest percentage of the total cost for both blood-banks (47.5% in Seccional-Bolivar and 55.7% in Yopal-Casanare). Conclusion: Our results are an important input for cost-effectiveness studies regarding screening tests for Chagas disease in Colombian blood-banks.