Abstract: The study aimed to: 1- Assess knowledge and practice of nurses regarding prevention of infection for children under hemodialysis. 2- Evaluate the effect of educational nursing program on nurses' knowledge and practice regarding prevention of infection among children undergoing hemodialysis. Research Design: Quasi experimental research design. Setting: The study was conducted in renal dialysis unit at Assiut Children University Hospital. Subjects: A convenient sample of 32 nurses working in the pediatric renal dialysis unit who accepted to participate in this study. Tools: 1- Structured questionnaire sheet about nurses knowledge including sociodemographic data; nurses' knowledge about renal failure, renal dialysis, general precautions and general precautions during nursing care for children under hemodialysis. 2- An observational checklist sheet: It was developed by the researcher to assess nurses' practice for children under hemodialysis. It was applied before and after 2 months of the implementation of the educational program. It consisted of - general and specific precautions during care of children under hemodialysis and application of infection control measures. Results: The levels of knowledge score were satisfactory among 90.6% of nurses. There was highly significant improvement of the nurses practice scores regarding general precautions, hand washing, wearing gloves, wearing mask, catheter insertion, skin preparation technique, care of patients equipment, dealing with sharp objects, dealing with blood, body secretions and fluids, maintain clean environment and safe injection practices in addition to the total practice score. There was significant improvement in items of the nurses' practice as laboratory-style covering coats, full isolation, and catheter care. The practice score level was adequate among 68.7% while inadequate among 31.3% of nurses. There were significantly higher nurses' knowledge score and levels of improvement of their knowledge score after the education program among older nurses and those with Bachelor of nursing. Conclusion: Nurses' knowledge and practices regarding hemodialysis and infection control guidelines are inadequate. Application of nursing education program about infection control measures when dealing with children under hemodialysis had significant improvement in nurse's knowledge and practice. Recommendations: Nurses should be encouraged to attend specific meetings as workshops and seminars held for infectious diseases and infection control to be acquainted with the recent advances and skills in hemodialysis unit.