The purpose of this study was to investigate the difference of fitness performance in collegiate level tennis players. Methods: seventy-one collegiate level tennis players (forty-nine males, twenty-two females) were recruited to participate in this study, which were divided into the elite group (E group, 40) normal group (N group, 31) according to their collegiate ranking. All participants had to finish eight fitness tests, including the flexibility (sit and reach, SR), muscular strength (throwing, TR), muscular endurance (sit-ups, SU), explosive power (standing long jump, SLJ), agility (ten meter shuttle run and x type shuttle run), speed (fifty meter sprint) and cardiorespiratory endurance (twelve minute running) tests. Two-way ANOVA was used to investigate the difference between in gender and the group's performance. Results: SLJ, TR, SU, ten meter shuttle run, x type shuttle run and twelve-minute running tests had significant differences in gender and group; the main effects analysis shown SLJ; Throwing and ten meter shuttle run tests had significant differences both in two groups and males are better than females, respectively. Skill performance in the female E group was better than N group. Conclusions: SLJ, TR, SU, ten meter shuttle run, x type shuttle run and twelve minute running tests shown significant differences in gender and group. We suggest that tennis periodization programs should focus on these special fitness performances.