We evaluated the sensitivity of the Yo-Yo intermittent recovery test-level 1 (Yo-Yo IR1) when discriminating among players in varying playing positions and different age categories in youth soccer. One-hundred and six prospective young soccer players, grouped on the basis of chronological age (under-13, under-14, under-15, under-16, under-17, under-18, and under-19) and playing position (center-backs, fullbacks, center midfielders, wide midfielders, and forwards), participated in the study. The players were administered a single Yo-Yo IR1 test at the beginning of the spring season. Analysis of variance revealed significant (F = 25.3; p < 0.001) group differences in Yo-Yo IR1 test performance scores among the observed age categories, and a systematic age-related increase in the Yo-Yo IR1 test performance was evident. Subsequent post hoc comparisons identified a number of significant differences among the selected age categories in Yo-Yo IR1 test performance. Analysis of covariance identified significant differences among playing positions (F = 3.1; p < 0.019) in the Yo-Yo IR1 test performance after controlling for age (F = 135.1; p < 0.001). Subsequent pairwise comparisons of the adjusted Yo-Yo IR1 test performance identified that center-backs had achieved significantly lower (all p < 0.01) performance scores than center midfielders, wide midfielders, and forwards, but not fullbacks. These results could be of practical value to coaches and scientists for further development of talent selection and profiling procedures in soccer, particularly because (a) the endurance performance represents a very important fitness component in selection and profiling of young soccer players and (b) the Yo-Yo IR1 test proved to be valid, reliable, and easily available measurement tool of a player's soccer-specific endurance capacity.