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Background: Hamstring muscles are most affected by multiple sprint-based sports as a result of muscle strain during sprinting, leading to reduced performance and increased risk of injury. Therefore, the purpose of the study was to assess inter-individual variability in hamstrings recovery after a sport-specific repeated-sprint training (RST), through sprint-specific markers of muscle recovery and associated muscle damage biomarkers in women and men. Methods: Healthy females ( n = 14) and males ( n = 15) underwent 10 repeated 40-m sprints with a 3-min rest pause between each repetition. Force-generating capacity (FGC) by the 90° hip :20° knee test and range of motion Jurdan test , together with serum biomarkers [sarcomeric mitochondrial creatine kinase (sMtCK), oxidative stress, irisin] were tested at baseline and 24-, 48- and 72-h post-exercise through a repeated measures design. Participants were classified according to FGC loss into high responders (HR) and low responders (LR). Results: 21 individuals (10 females, 11 males) were classified as HR (FGC loss >20% and recovery >48 h), while 8 individuals (4 females, 4 males) were classified as LR. HR individuals showed unrecovered maximal voluntary isometric contraction (MVIC) torque until 72 h post-training ( p = 0.003, n p 2 = 0.170), whereas only HR males showed decreased range of motion ( p = 0.026, n p 2 = 0.116). HR individuals also showed increased sMtCK ( p = 0.016, n p 2 = 0.128), oxidative stress ( p = 0.038, n p 2 = 0.106) and irisin ( p = 0.019, n p 2 = 0.123). Conclusion: There is inter-individual variability in the muscular response to a sport-specific RST, identifiable by MVIC torque assessment. The findings support that the 90° hip :20° knee test is a powerful indirect test to screen hamstrings recovery in both women and men, in a cost-effective way. However, the Jurdan test might not be able to monitor hamstrings recovery in sportswomen after RST. Decreases in muscle capacity are linked to damage to muscle sarcolemma and mitochondria until 72 h post-exercise. Overall, 72 h will not be adequate time to restore hamstrings structure and function after a sport-specific RST in both female and male responders., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer JT declared a shared affiliation with the authors to the handling editor at the time of review., (Copyright © 2024 Cosio, Moreno-Simonet, Porcelli, Lloret, Padulles, Padulles, Farran-Codina and Cadefau.)