Background:The participation number of nurses hand hygiene was low. It caused high number of Health-care Associated Infections (HAIs). The high number of HAIs caused time of patients treatment much longer, mortality, increased treatment fee and made treatment will be more dificult. Therefore, identification of hand hygiene implementation in the hospital would be the first step to give intervention to increase the nurses participation. The goal of this research is to identify hand hygiene implementation of the nurses in RS PKU Muhammadiyah Yogyakarta Unit 2. Method:The research used analytic descriptive methode. The samples of the research were all nurses of RS PKU Muhammadiyah Yogyakarta Unit 2 who work at education ward and 110 act of hand wash. Data analyzing of the research used frequent distribution. The result of the research showed that the mayority of the nurses did not obey doing HH base on the SPO RS PKU Muhammadiyah Unit 2. Result:The implementation of the 5 moment was done most in 4 moment. The majority of the nurses did HH incorrectly base on the six steps procedure of HH. They did HH incorrect time base on the HH duration from WHO. The nurses level of knowledge of hand hygiene was in enough level, 11 nurses. Conclussion: The findings suggest that implementation of nurses hand hygiene must be improved under supervision by hospital management. Keywords: hand hygiene, nurses.