This study aimed to examine the effect of the hand hygiene education on middle school students' behavioral scores related to hand hygiene practices. The research adheres to a quasi-experimental design suitable for the pre-test Posttest model with a control group. The population of the study consists of a total of 1159 students, comprising 5th, 6th, 7th, and 8th graders. During the first semester of the 2021-2022 academic year, face-to-face education was conducted at a middle school located in Ankara. Two face-to-face training sessions were conducted to increase awareness of hand hygiene. The groups indicate homogeneity in terms of demographic characteristics. The mean hand hygiene behavior score was 56.45 ± 8.03 before the education in the intervention group, which increased to 59.79 ± 7.62 after the education. In the control group, the mean score was 55.98 ± 7.89 before the education and 57.15 ± 7.63 after the education. It was found that after the educational program, there was a statistically significant increase in hand hygiene behavior scores of the groups. The research findings indicate a beneficial impact of hand hygiene education on students' average behavioral scores. To foster lasting behavioral changes from a young age, it is advisable to implement hygiene education in schools periodically, including targeted interventions for middle school students.