Little is known about the motivational factors or design features that might lead female career and technical education (CTE) health sciences students to persist in self-regulated engagement to win state achievement in serious simulation games (SSGs). In this study, 12th grade female CTE health sciences students played a 3-Level hand hygiene SSG that was developed based on a learner analysis of a prior cohort of students. A within-subjects repeated measures design was used to assess differences in student motivation using a 10-construct Value-Achievement-Cost (VAC) Survey between baseline and the time they exited the SSG. Student choice to persistently engage in each level of the SSG was also evaluated using a 10-item SSG design feature score board. The average time that the Level III achievement students were engaged in the SSG was twice as long as Level I/II achievement students. Three of the 4 perceived mean Cost construct scores significantly decreased with large effect from baseline for students who persisted to the SSG Level III win state while all pre-post Value and Achievement orientation constructs scores remained unchanged. Mean average differences were not statistically significant for any of the ten core SSG design elements regardless of engagement time.