We examined the validity and reproducibility of a squash-specific multiple-sprint test. Eight male squash and 8 male soccer players performed Baker's 8 × 40-m sprints and a squash-specific-multiple-sprint test on separate days. The sum of individual sprint times in each test was recorded. Six squash and 6 soccer players repeated the tests 7 days later to assess reproducibility using intraclass correlation. In addition, 2 England Squash coaches independently ranked the squash players using knowledge of the player and recent performances in local leagues. Performance on the squash-specific (r = 0.97 and 0.90) and Baker's test (r = 0.95 and 0.83) was reproducible in squash and soccer players, respectively, and did not differ on Baker's test (mean ± SD 72.9 ± 3.9 and 72.9 ± 2.8 seconds for squash and soccer players, p = 0.969, effect size = 0.03). Squash players (232 ± 32 seconds) outperformed soccer players (264 ± 14 seconds) on the squash-specific test (p = 0.02, effect size = 1.39). Performance on Baker's and the squash-specific test were related in squash players (r = 0.98, p <0.001) but not in soccer players (r = −0.08, p = 0.87). Squash-player rank correlated with performance on the squash-specific (ρ = 0.79, p = 0.02) but not the Baker's test (ρ = 0.55, p = 0.16). The squash-specific test discriminated between groups with similar non-sport-specific multiple-sprint ability and in squash players. In conjunction with the relationship between test performances, the results suggest that the squash-specific test is a valid and reproducible measure of multiple-sprint ability in squash players and could be used for assessing and tracking training-induced changes in multiple-sprint ability.