Abstract. INTRODUCTION: The most important tool in any infection control program is good hand hygiene. Despite recognizing that hand hygiene is crucial in reducing infection rates, hand hygiene compliance remains suboptimal. This study was designed to determine hand hygiene compliance, before and after an educational intervention and its impact on hospital infection rates. METHOD: The study was done in neonatal intensive care unit of an educational hospital. All healthcare providers working in the unit at the time of study were trained on importance of hand hygiene and methods of hand hygiene observation; after that hand washing compliance controlled by a physician during postintervention phase. Hand hygiene compliance, healthcare associated infection and mortality rates compared before and after educational intervention. RESULTS: Compliance of health-care workers for all hand hygiene opportunities combined was 30% before intervention and improved to 70% in postintervention. In postintervention phase, healthcare associated infection rates and mortality rates decreased significantly as the hand hygiene compliance improved. CONCLUSIONS: Good control of hand hygiene compliance by physician after an educational program may have good effect in healthcare associated infections control in neonatal intensive care unit.