Background: Evidence-based nursing (EBN) is the essential of quality care. To ensure evidence-based practice relies on the nurse s competence. However, little study reports the content and effectiveness of the EBN training program. Purpose: The purpose of this study was to evaluate the effectiveness of an EBN training program on nurses knowledge, attitude, and behavior. Methods: The study was a pretest-posttest research design. Seventy-two nurses received an EBN training program. The program includes: (a) 18 hours lectures, including concepts and steps of EBN, sources and search of evidences, critical appraisal, evidence-based practice and guideline development, teaching in EBN, (b) 10 weeks small group practicum and mentoring, and (c) 9 hours presentation and sharing. The EBN knowledge, attitude, and behavior were measured using a questionnaire before and after implementing the training program. The Paired-t statistics was used for analyzing the effectiveness of the training program. Results: There was no significant difference of EBN knowledge and attitude scores between pretest and posttest. However, the posttest score of EBN behavior was significantly higher than the pretest (p & .001). Conclusions: The EBN training program improves the evidence-based nursing behavior. The content of this program can be a reference for designing a systemic training program of other institutes.