Background: Non-tunneled hemodialysis catheters are typically used when vascular access is required for urgent renal replacement therapy Nurses play a crucial role in tracking the occurrence of complications such as catheter exit site infection, thrombosis, and bleeding. Aim: This study aimed to evaluate the effect of guidelines on nurses' performance regarding care of patient with non-tunneled hemodialysis catheter. Research design: Quasi-experimental design study was conducted to achieve the aim of the study. Setting: This study was conducted in the dialysis unit at Benha University Hospital. Sample:  A Convenient sample of (58) nurses working in dialysis unit during the period of the study. Tools: Two tools were used in this study, Tool 1: Structured questionnaire to assess nurses' knowledge regarding care of patient with non-tunneled hemodialysis catheter. Tool 2: Included nurses' practice observational checklist toward care of patient with non-tunneled hemodialysis catheter. Results: 6.9% of the studied nurses respectively have good level of total knowledge regarding care of patient with non-tunneled hemodialysis catheter pre implementation of educational guidelines. While improved to 89.7% post implementation and slightly decreased to 84% & 86.7% at follow-up phase. 82.8% of the studied nurses respectively were satisfactory in the use of the Observational Tool pre implementation of educational guidelines. While improved to 86.2% post implementation and slightly decreased to 84% at follow-up phase. There was a highly significant statistical positive correlation between nurses' knowledge and practice in Benha University Hospital at phases of educational guidelines implementation at p < 0.01. Conclusion: Implementation of educational guidelines had statistically significant improvement of nurses' knowledge, and practice regarding care of patient with non-tunneled hemodialysis catheter. Recommendations: The present study recommended that continues education and in service training program for nurses to improve their performance regarding care of patient with non-tunneled hemodialysis catheter.