Background: Infectious diseases usually cause morbidity and mortality all over the world. However, hand-washing practice is the most important methods to prevent the transmission of infectious diseases. Aim: Therefore, this study aimed at assessing the effect of hand-washing education on knowledge and practice of infectious diseases prevention in selected health centers in Akure South, Ondo State. Methodology: The study employed a pre-test and post-test design, using questionnaires to obtain data from 98 women in selected health centers in Akure South, Ondo State. Data were analyzed using SPSS 20.0. Descriptive statistics such as frequencies, percentages and chart were used to summarize and present data. Furthermore, associations between variables were tested using ANOVA at p value 0.05 level of significance. Results: The result from this study revealed better knowledge of hand-washing and good understanding of hand-washing practices after hand-washing education. Frequent education on hand hygiene and importance of hand washing, notice to create awareness on hand-washing at every corners of health centres, availability of running water and liquid soap at various locations of the clinic were observed measures put in place to ensure the strict adherence to hand-washing among patients. The researcher found out that there was a significance difference between hand-washing knowledge and practices before and after hand-washing education (P = 0.00, P < 0.05). Conclusion: It is therefore concluded that hand-washing education improves the respondents knowledge on hand-washing and hand-washing practices.