Purpose: This study was conducted to develop and test the effects of a program for practice application of intravenous infusion evidence based nursing practice (EBP) guidelines in small and medium-sized hospitals. Methods: A mixed method research design was used, combining non-equivalent control group pre-post test design with qualitative study analysis. The subjects consisted of 55 nurses. The practice application program was developed based on the Advancing Research and Clinical practice through close Collaboration (ARCC) model. Data were collected for analysis in the following areas: nurses’ EBP organizational culture and readiness, EBP beliefs, EBP implementation, importance about intravenous infusion, and performance about intravenous infusion, with data assessed using valid and reliable instruments. Patient outcomes were collected from the hospital’s medical records. Data were analyzed using t-test, χ2-test, and Shapiro-Wilk test, with qualitative content analysis used for interview data. Results: Following the intervention, nurses’ EBP organizational culture and readiness, EBP beliefs, EBP implementation, and performance of intravenous infusion and perceptions of its importance showed significant improvement in the experimental group. Phlebitis rates decreased in the experimental group compared to the control group. Conclusion: This program is effective to improve nurse’s perception and practice of evidence-based nursing. Therefore we recommend to use this program at same levels of hospitals.