Background of the Problem: The prevalence of TB in Indonesia ranks 2nd in the World; therefore efforts to prevent TB transmission are needed, including through the use of PPE in high-risk individuals. This study aims to determine the relationship between knowledge and behavior of nurses in the use of PPE to prevent TB transmission in hospitals. Research Methods: This study used a cross-sectional design. The subjects in this study were 38 respondents, with a total sampling technique. The instrument used was a questionnaire — data analysis using descriptive statistical test and chi-square test. Research Results: The results of this study showed that most nurses had good knowledge n = 31 (81.6%). Most nurses showed good behavior n = 32 (84.2%). This study also showed that there was a significant relationship (p-value 0,0006) of knowledge with nurse behavior in the use of PPE for the prevention of TB transmission. Conclusion: There is a significant relationship between knowledge of nurse behavior in the use of PPE for the prevention of TB transmission.