Abstract Introduction: Educational program about central venous catheter bundle plays an important role on critical care nurses' knowledge and practice. Educating, and training health care providers who are inserting and maintaining central venous catheter is must. Aim: To evaluate the effect of educational program on critical nurses' knowledge and practice about central venous catheter bundle. Subjects and Method: Quasi-experimental research design was used. The study executed at General ICU (18 beds), at main Assiut University Hospital. A sample of 60 critical care nurses' working in General ICU was selected. Two tools were used for this study: knowledge assessment tool, and practice assessment tool. Results: There was a significant increase in the percentage of satisfactory knowledge and practice of critical care nurses after educational program compared with that before educational program (86.7% and 100% Vs. 0.00% and 15%) for knowledge and practice, respectively with minimal decline of satisfactory knowledge at follow up to 75 % and 81.7% for knowledge and practice, respectively. Conclusion: A significant improvement in the knowledge and practice of critical care nurses regarding central venous catheter after educational program was noted. Recommendation: Training programs for critical care nurses to keep up with updated knowledge and evidence practice.