Background: Nosocomial infections are a significant problem in paediatric intensive care units (ICUs). The hand hygiene of health-care professionals is the main measure for preventing hospital infections. Aim: Assessment of the effectiveness of edu- cational intervention on the compliance of health professionals with the recommendations for hand hygiene and investigation of the opinions of health-care professionals about nosocomial infections and educational intervention. Method: A survey was conducted of nurses, doctors and physiotherapists who work in the ICU of a paediatric hospital in Athens. The collection and analysis of data was based on the “Observation and Calculation Form − World Health Organization: Hand Hygiene Technical Reference Manual” of the World Health Organization. The questionnaires “Perception Survey for Health-Care Workers” and “Follow-up Perception Survey for Health-Care Workers” were completed by the health-care workers to assess their perceptions of health care-associated infection and hand hygiene before and after the educational programme, respectively. The test reli- ability was assessed using the Cronbach’s alpha, and was 0.81 before and 0.89 after the programme, respectively. Statistical analysis was conducted using the Statistical Package for Social Sciences (SPSS) v. 20.0 included x2 test, with a significance level of p<0.05. Results: The total numbers of observations studied were 434 before and 409 after the programme. The degree of compliance of health professionals with the recommendations for hand hygiene before educational intervention was 31.8% and after 51.5% (x2=33.2, p<0.001). Among doctors, nurses and physiotherapists compliance increased from 28.3%, 30.4% and 36.2% before, to 40.3%, 54.2% and 36.3% after intervention, respectively. The majority of health-care workers believed that the educational activities that they participated in were important to improve their hand hygiene practices and that the results of hand hygiene observation in their ward helped them to improve their hand hygiene practices. Conclusions: The degree of compliance of the staff with recommendations for hand hygiene in the ICU increased adequately after educational intervention. The training and information of health professionals contributes to increasing the degree of compliance with the international recommendations for hand hygiene.