Background: Hospital as a health service institution should be able to prevent the risk of infection for patient and hospital workers. One of indicators success in hospital are the low of nosocomial infections in hospital.Cleanliness of infrastructuresin hospital is an attempt to prevent nosocomial infection , so that the Cleaning Staffs’s knowledge about cleanliness of facilities and infrastructure is very important. Based on Diskes RI data in 2004, the proportion of the data nosocomial infections 55,1 % happened in government hospital, while 35,7 % happened in private hospitals and 9,1 % going on with the hospital in ABRI. Purpose: The purpose is to know the effect of education about sanitation of hospital’s facilities in increasing knowledge of PKU Muhammadiyah Gamping hospital's cleaning staffs. Method: The method for this research is a quantitative research with quasy experiment pretest-posttest control group design. The sample is cleaning Staffs in PKU Muhammadiyah Gamping while the respondents are 24 of it. They are collected by a total sampling technique. Data were analyzed by Paired SampleTTest to know the influence of education between before and after education in intervention and control group and Independent Sample T Test to know the mean of knowledge between intervention and control group. Result and discussion: The result for this research confirmed that there are 24 Cleaning Staffs have the same level of knowledge on the value of pretest. The result proved in different tests that show p value <0,05. The results showed that the intervention group have the increasing of knowledge abaout hygiene infrastructure hospital and the control did not have the increasing of knowledge abaout hygiene infrastructure hospital. The value of P in the intervention group (p=0,003) and the value of P in the control group (p=0,651). So, education can increase the knowledge of Cleaning Staffs. Conclusion: Education about sanitation of hospital’s facilities can effect the knowledge of of PKU Muhammadiyah Gamping hospital's cleaning staffs.