ObjectiveTo investigate hand hygiene compliance of health care workers(HCWs)  in a tertiary hospital, and to explore effective methods to improve hand hygiene compliance.MethodsBy direct observation method, hand hygiene compliance and practice among HCWs in 23 clinical departments were surveyed in SeptemberDecember 2012, and hand hygiene compliance monitor forms were filled in. ResultsA total of 721 opportunities of hand hygiene were recorded, overall compliance rate of hand hygiene was 58.53%, 32.18% used instant hand antiseptic, 26.35% selected running water, and 19.42% wore gloves without taking hand hygiene measures. For different occupations, hand hygiene compliance among nurses was the highest (68.65%), and cleaners was the lowest (39.25%). Of the five moments for hand hygiene, hand hygiene compliance after body fluid exposure was the highest (73.75%), followed by the moment after touching a patient (65.00%), while the moment after touching patients’ surroundings was the lowest (47.85%). ConclusionHand hygiene compliance of HCWs needs to be improved, and the management should be further strengthened, corresponding measures should be taken for different personnel.