The purpose of this study was to design and evaluate a STD prevention education program which was developed based on Health and Nursing Curriculum Guidelines of High School. This study adopted nonequivalent experiment control group design. A total 1023 students from 14 senior high and vocational schools in Taiwan participated in the study. The intervention group (N = 535) received a 9 hours STD prevention education program which developed based on the sprit of the holistic sexuality education. The control group (N = 488) received ordinary classroom sessions of Health and Nursing. The teachers of intervention group were trained to implement this program. The outcomes of intervention were measured by questionnaires administered on week before (pre-test) and after (post-test) the intervention. The results indicated that the target of enhancing core abilities in STD prevention can be better achieved by the STD prevention education program which setting in the Health and Nursing Curriculum Guidelines of High School than by ordinary sexual education while increasing no added loads to teachers. The program had statistically significant positive impact on the knowledge of “holistic sexuality”, “safer sexual behavior”, “the health of birth and abortion”, “AIDS prevention”, “the prevention of sexual harassment and assault”; the attitude of “the comprehension of homosexual”, “safer sexual behavior”, “AIDS prevention”; the self efficacy of “skills of healthy interaction between both sexes”, “safer sexual behavior”, “the prevention of sexual harassment and assault”, and behavior intention of “safer sexual behavior”. Suggestions were made in terms of education and research in the future according the results of the study.