This study was conducted to develop and evaluate a web-based infection prevention education program for children, parents and teachers. Research for development of the web-based education program was completed in four phases (analysis, design, development, and evaluation) from 1 February 2015 to 5 October 2015, and the completed website was named CHILD4HEALTH (http://uwcms.pusan.ac.kr). Educational contents pertaining to infection prevention were composed of three sections, children, parents and teachers. Subjects were divided into nine categories, animation, children's dictionary, with mom, music, games, quizzes, educational contents for parents, educational contents for teachers, school newsletters, and handouts. Six characters were developed to increase interest and educational effect. Program evaluation items comprised the website, reliability, and satisfaction. Website evaluation by parents revealed that ease of use was $3.77{\pm}0.70$, entertainment value was $4.07{\pm}0.27$, childproof was $3.82{\pm}0.67$, education value was $4.02{\pm}0.75$, and design features were rated $3.65{\pm}0.53$. According to teachers, ease of use was $3.98{\pm}0.37$, entertainment value was $4.00{\pm}0.17$, childproof was $4.34{\pm}0.60$, education value was $4.00{\pm}0.20$, and design features were $3.81{\pm}0.56$. Parents scored reliability and satisfaction as $8.33{\pm}0.62$ and $7.80{\pm}0.77$, respectively, while they were scored as $8.50{\pm}0.73$ and $8.10{\pm}0.74$ by teachers. Based on the results of this study, the developed web-based education program will help prevent infectious disease and facilitate development of future education programs regarding such diseases.