Objective: The serological diagnosis of syphilis has a range of false positives varying from 1 to 3%. The use of treponemal and nontreponemal tests for screening as well as the implementation of automatic methods has been changing constantly in an effort to improve sensitivity and specificity parameters. We assessed the sensitivity for the syphilis screening of treponemal and nontreponemal tests in blood donors at Hospital Pablo Tobón Uribe. Methodology: An observational, cross-sectional study conducted from January 2012 to December 2014. Results from 342 patients were registered with positive serology by treponemal and nontreponemal tests, using the FTA-ABS as gold standard in the blood donors scenario. Results: The sensitivity percentages of the nontreponemal test RPR and the automatic test Architect were 29.3% (CI 95% 22.9-35.6) and 99.53% (CI 95% 98.39-100), respectively. Conclusion: the Architect treponemal test has a better sensitivity than the nontreponemal test RPR. In the scenario of syphilis screening in blood donors, the treponemal test reduces the mistakes when detecting the infectious disease.