The purpose of this study was to assess the validity of the recently developed Basketball Exercise Simulation Test (BEST). Ten semiprofessional (age, 22.7 ± 6.1 years; height, 189.6 ± 9.5 cm; weight, 86.5 ± 18.7 kg; % body fat, 14.7 ± 3.5%) and 10 recreational (age, 26.6 ± 4.0 years; height, 185.9 ± 7.9 cm; weight, 92.6 ± 8.4 kg; % body fat, 23.8 ± 6.3%) male basketball players volunteered for the study. The participants completed a Yo-Yo Intermittent Recovery Test (Yo-Yo IRT) and BEST trial midway through the playing season. Eight participants (semiprofessional, n=4; recreational, n = 4) completed an additional Yo-Yo IRT and BEST trial at the end of the playing season. Performance measures from the BEST included sprint decrement (%), mean sprint and circuit time (seconds), and total distance covered (m). Construct validity was calculated using Student's unpaired t-tests to identify the differences in Yo-Yo IRT and BEST performances between playing levels. Longitudinal validity was determined based on the relationship between changes (%) in Yo-Yo IRT1 and BEST performances across the season. Semiprofessional players performed significantly (p, 0.01) better in the Yo-Yo IRT (1,283±62 vs. 636±297 m) and BEST (mean sprint time: 1.45 ± 0.01 vs. 1.65 ± 0.03 seconds; mean circuit time: 18.98 ± 1.79 vs. 22.72 ± 2.01 seconds; sprint decrement: 8.54 ± 0.15 vs. 15.38 ± 0.27%) compared with recreational players. For the group as a whole, a strong relationship was evident between the changes in BEST sprint decrement and changes in Yo-Yo IRT performance (R =20.815, p = 0.014) across the season. In conclusion, the BEST displayed both discriminative and longitudinal validities and provides a novel match-specific fitness test for basketball players.