Purpose: The purpose of this research was to identify the relationships between stroke knowledge, health perception, exercise self-efficacy and stroke prevention behaviour and the factors influencing stroke prevention behaviour in middle-aged adults. Methods: A cross-sectional survey was conducted in 2 cities of Korea from May to July 2020, using structured questionnaire. The participants were 168 middle-aged adults without a history of stroke. Collected data were analyzed using descriptive statistics, independent t-test, one-way ANOVA, Pearson correlation coefficients, and multiple regression with SPSS/WIN 25.0. Results: There was significant correlations among degree of stroke prevention behaviour, stroke knowledge about warning sign (r=.20, p=.010), health perception (r=.35, p<.001) and exercise self-efficacy (r=.43, p<.001). The most important factor influencing stroke prevention behaviour was exercise self-efficacy (β=0.38, p<.001), followed by health perception (β=0.18, p=.008), body mass index (β=-0.17, p=.011), stroke knowledge about warning sign (β=0.13, p=.045) in that order. These factors explained 37.7% of total variance in stroke prevention behaviour (F=11.09, p<.001). Conclusion: The results of this study suggest that the development of nursing intervention for stroke prevention behaviour improvement is needed considering exercise self-efficacy and stroke knowledge.