We compared knowledge and self-reported hand hygiene practices with direct observations among Shiraz Nemazee Hospital nurses. Overall hand hygiene compliance was low (39.6%). Use and appreciation for alcohol-based handrubs were suboptimal. Some reluctance was due to religious concerns. Most nurses self-reported performing proper hand hygiene more than 75% of the time, whereas direct observation indicated much lower levels of compliance. It appears that additional training and improved monitoring are needed.