Based on recommendations from WHO, hand hygiene (HH) is the most important way to control hospital infections. Due to the critical role of nurses in pediatric intensive care units (PICUs), they should have essential and updated information regarding HH. Aim: This study was aimed to investigate nurses’ compliance with hand hygiene (HH) and its relation to health care acquired infections (HCAIs) at PICUs. Setting: the current study was conducted in Neonatal and Pediatric ICUs at Benha Specialized Pediatric Hospital. Design: A descriptive design was utilized. Sample: A convenient sample of 100 nurses and 414 pediatric patients were included. Tools: Tool one; an interviewing questionnaire schedule to evaluate nurses’ general characteristics, their knowledge towards compliance with hand hygiene and finally barriers regarding hand hygiene. Tool two; an observational checklist to assess nurses’ practical compliance with hand hygiene. Results: It was evident from the present study results that the mean age of the studied nurses was 29.03 ± 9.70 years and more than one third of them had more than five years of experience in PICUs. Regarding knowledge, 72% of nurses had incorrect knowledge about HH compliance, more than two thirds (67%) of them had incompetent practice regarding HH procedure and most of the studied nurses (81%) weren’t comply with HH. In addition, there was a highly statistical significant difference (P |