The present study investigated the test-retest reliability from the Yo-Yo IR1 (distance and heart rate responses), and the ability of the Yo-Yo IR1 to differentiate between elite and non-elite youth soccer players. A total of 228 youth soccer players (11-17 years) participated: 78 non-elite players to examine the test-retest reliability within 1 week, added with 150 elite players to investigate the construct validity. The main finding was that the distance covered was adequately reproducible in the youngest age groups (U13 and U15) and highly reproducible in the oldest age group (U17). Also, the physiological responses were highly reproducible in all age groups. Moreover, the Yo-Yo IR1 test had a high-discriminative ability to distinguish between elite and non-elite young soccer players. Furthermore, age-related standards for the Yo-Yo IR1 established for elite and non-elite groups in this study may be used for comparison of other young soccer players.