Background: Ventilator associated pneumonia is the most common hospital-acquired infection among patients receiving mechanical ventilation in pediatric intensive care unit. Using an evidence based guidelines for the prevention of ventilator associated pneumonia (VAP) are recommended. Objectives: to investigate nurses' knowledge and practice toward evidence-based guidelines for the prevention of ventilator associated pneumonia in pediatric intensive care units Methods: Aquasi- experimental pre-post intervention study was done on 51 nurses of different categories working in medical, neurology and surgical ICU at Mansoura University children's Hospital, Egypt. Three tools were used; An interview questionnaire sheetwas developed by the researcher in Arabic language. Nurses' knowledge of evidence-based guidelines for the prevention of VAP, multiple choice questionnaire and observational checklist for nurses' performance to ventilator bundles for VAP prevention. Results:  mean score level of nurses general knowledge about VAP, Knowledge of Evidence-based guidelines for VAP prevention and nurses performance to ventilator bundled for VAP prevention where the mean score were 8.6078±1.9908, 2.4706±1.3469, 2.1400±1.4287 respectively before program implementation compared to 12.1569±1.0464, 7.2941±1.7005, 5.5800±1.5265 respectively after program. There is a highly significant difference between nurses knowledge and practices to Evidence-based guidelines for prevention of VAP before and after the program (t= -11.059, -22.106, -13.796 and p = .000, 0.000, 0.000 respectively). Conclusion:  Adequate improvement in nurses' knowledge and practice after applied evidence based guidelines for prevention of ventilator associated pneumonia in pediatric intensive care unit.