Background: Umbilical Venous Catheterization (UVC) is a life-saving procedure at neonatal intensive care units in neonates who require vascular access and resuscitation. The nurse has a vital role in increasing the usable life span and minimizing the catheter associated complications. Aim: This study aimed to evaluate the effect of nursing interventions on nurses' performance regarding care provided to neonates undergoing umbilical venous catheter.   Design: A quasi-experimental design (pretest and posttest) was utilized in this study. Settings: The study was carried out at Neonatal Intensive Care Units (NICU) of Behna University Hospital and Benha Specialized Pediatric Hospital Sample: A convenience sample of all available bedside/ working nurses (n= 90) and a purposive sample of (90) neonates admitted to NICU at the time of the study Tools of data collection: Two tools were used, I: Socio-demographic characteristic and structured interviewing schedule including three parts: a) Nurses socio demographic characteristics, b) Neonates characteristics and medical history and c) Nurses' knowledge regarding care provided to neonates undergoing UVC and II: Observational checklists for nurse's performance regarding care of neonates undergoing UVC. Results: Knowledge and performance of nurses were higher on post (91.1% Vs 41.1% and 86.7% Vs 32.2%) than pre guidelines. There was a positive correlation between the studied nurses' total knowledge and practices scores regarding care of neonates undergoing UVC on posttest. Conclusion: The program had a positive effect in improving nurses' knowledge and practices post program implementation.Recommendation: Regular and continuous educational programs should be developed for nurses in NICU according to their needs aiming to upgrade their knowledge and improve their practice regarding to care of neonates undergoing umbilical venous catheter.