Background: Urinary tract infection is the most common healthcare associated infection (HCAI) that usually results from prolonged use of urinary catheter that results in prolonged hospital stay. Prevention of catheter associated urinary tract infection (CAUTI) in pediatric patients regarded now as an important hospital policy worldwide. Aim: The aim of this study was examining the effectiveness of evidence-based guidelines on catheter associated urinary tract infection rate among pediatric intensive care children. Design: A quasi-experimental design was utilized. Setting: The study was conducted in acute care units (medical pediatric intensive care unit “PICU", surgical PICU and cardiac PICU) at Mansoura University Children's Hospital. Subjects: The subjects of the study included 84 nurses who are working at the above-mentioned study settings &102 pediatric patients who are divided randomly into 2 identical groups, case& control groups, each group consists of (51) pediatric patients. Tools: I: A Structured self-administered questionnaire, II: evidence-based guidelines on Urinary catheter maintenance and removal observational checklists, III: A structured questionnaire of pediatric patients' health condition & IV: Catheter-associated urinary tract infection criteria checklist. Results: There was an improvement in the mean scores of nurses' total knowledge and practice with statistically significant difference (P<0.001) in the post intervention compared to pre intervention stage, additionally, the positive symptoms and signs suggesting CAUTI were more evident in control group compared to case group with significant difference (P <0.05 & P <0.001) post intervention. Conclusion: the implementation of evidence-based guidelines about CAUTI prevention for pediatric patients with urinary catheter has positive significant improvement on the prevention of CAUTI rate. Recommendations: there is a requisite to educate the pediatric nurse to avoid routinely use of urinary catheters and remove them when they are no longer required. The use of urinary catheter evidence-based guidelines checklists should be kept in the pediatric patient file and reviewed regularly by pediatric nurses.