To evaluate infection control and hand hygiene understanding at 3 public hospitals, we surveyed 4,345 healthcare workers (HCWs) 3 times during a 5-year infection control intervention. The preference for the use of alcohol hand rub for hand hygiene increased dramatically; in nurses, it increased from 14% to 34%; in physicians, 4.3% to 51%; and in allied HCWs, 12% to 44%. Study year, infection control interactive education-session attendance, infection control knowledge, and being a physician or allied HCW independently predicted a preference for alcohol hand rub.