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The purpose of this study was to investigate the association of acceleration and changes in spatiotemporal variables at each step with 100-m sprint performance in preadolescent sprinters. Twenty-six boys performed 100-m sprints, and their spatiotemporal variables were measured at each step. Acceleration was negatively correlated with the 100-m sprint time from the 1st to 21st step. The rates of change in step frequency were positively correlated with acceleration at the 2nd and 3rd step. Posterior to 3rd step, rates of change in step length were positively correlated with acceleration. The results suggest that the acceleration caused by increase in step frequency and step length up to reaching to the maximal sprint velocity is effective for improving the 100-m sprint time.