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The aim of this study was to investigate the effect of karate specialization on the reaction time in specific karate kumite situations. In order to maximize the ecological validity of experimental set-up, we used our recently proposed and evaluated video-based method (Mudric et al., 2015). Within this method, the reaction time is calculated from the kinematic parameters recorded on both offensive action stimuli and defensive action responses. The results obtained from three groups of subjects (i.e., karate kumite, karate kata and beginners) indicate significant differences in reaction time between the beginners and both specialized karate groups. There were also prominent but not significant differences between the karate kumite and karate kata. These findings suggest that particular sport specialization could improve reaction time of an athlete in specific sport situations.