In this intervention study hand hygiene compliance and its influence on the spread of healthcare-associated infections were investigated. Through a hand hygiene education program the compliance improved from 42% to 55% on the investigated neonatal unit. In parallel the consumption of hand rub increased by >20 l per 1 000 patient days and the infection risk of very low birth weight neonates was reduced. Interestingly, bacteremia by clonally related pathogens, indicating that pathogens were transmitted between neonates, were reduced as well.