Purpose: Hamstring muscle architecture may be associated with sprint performance and the risk of sustaining a muscle injury, both of which increase during puberty. In this study, we investigated the m. biceps femoris long head (BFlh) cross-sectional area (ACSA), fascicle length (FL) and pennation angle (PA), and sprint performance as well as their relationship in under 13 to 15 youth soccer players. Methods: We measured 85 players in under-13 (n = 29, age = 12.5 [0.1] y, height = 155.3 [6.2] cm, weight = 43.9 [7.6] kg), under-14 (n = 25, age = 13.5 [0.3] y, height = 160.6 [7.7] cm, weight = 47.0 [6.8] kg), and under-15 (n = 31, age = 14.4 [0.3] y, height = 170.0 [7.7] cm, weight = 58.1 [8.8] kg) teams. We used ultrasound to measure BFlh ACSA, FL and PA, and sprint tests to assess 10- and 30-m sprint time, maximal velocity  (vmax), and maximal acceleration (αmax). We calculated Pearson r to assess the relationship between sprint ability and architectural parameters. Results: All muscle architectural parameters increased from the under-13 to the under-15 age group (BFlh ACSA = 37%, BFlh FL = 11%, BFlh PA = 8%). All sprint performance parameters improved from the under-13 to under-15 age categories (30-m time = 7%, 10-m time = 4%, vmax = 9%, αmax = 7%). The BFlh ACSA was correlated with 30-m sprint time (r = −.61 (95% compatibility interval [CI] [−.73, −.45]) and vmax (r = .61, 95% CI [.45, .72]). A combination of BFlh ACSA and age best predicted 30-m time (R² = .47 [.33, .62]) and 10-m time (R² = .23 [.08, .38]). Conclusions: Muscle architectural as well as sprint performance parameters increase from the under-13 to under-15 age groups. Even though we found correlations for all assessed architectural parameters, BFlh ACSA was best related to the assessed sprint parameters.