This study investigated the feasibility and reliability of a 12 × 25-m repeated sprint test with sprints starting every 25-s in children aged 6–8 years (36 boys, 41 girls). In all subjects, total sprint time (TST) demonstrated high test-retest reliability (ICC: r = .98; CV: 0.7% (95% CI: 0.6–0.9)). While sprint time varied over the 12 sprints in all subjects (p < .001) with a significant increase in time for the third effort onwards compared with the first sprint (p < .001), there was no difference in performance between genders. In all subjects, TST decreased with age (p < .001) and was accompanied by an increase in estimated anaerobic power (p < .001) but also in sprint time decrement percentage (p < .001). Gender did not effect these changes. The present study demonstrates the practicability and reliability of a repeated sprint test with respect to age and gender in young children.