Hospital provides services with positive and negative impact. One of the negative impacts that could be happening is Hospital-Acquired Infections (HAIs). Personal Protective Equipment (PPE) is one of the way to reduce the incidence of HAIs, so that hospital employee’s basic knowledge about PPE is important. Based on World Health Organization (WHO) survey in 2011, HAIs prevalence in Indonesia (which is a medium-income country) is about 7,1%. Purpose: To analyze the relations between knowledge and Personal Protective Equipment (PPE) use adherence in non-medical employee at PKU Muhammadiyah Gamping Hospital. Methods: This is a quantitative with analytic-observational design and cross-sectional approach’s research. The sample is non-medical employees at PKU Muhammadiyah Gamping Hospital while the respondents are 19 of it. They are collected by a total sampling technique. Fisher’s Exact Test and Spearman Correlation Test are used to analyze the relations between both variables. Results and Discussion: This research confirmed that there are 16 employees with good level of knowledge (82,4%), 15 employees obeys the rule in using PPE (78,9%). Employee’s level of knowledge is highly related with the adherence in using PPE with p value = 0,004. The high level of non-medical employee’s knowledge is related to the level of their adherence with p value = 0,003 (Correlation coefficient = 0,651). Conclusion: There is a relation between knowledge and adherence in using PPE in non-medical employees at PKU Muhammadiyah Gamping Hospital and the higher the knowledge, the higher the adherence also. The correlation coefficient is about 0,651 which means the relation is tight