This retrospective epidemiological study investigated the relationship between physical fitness testing and locomotive disorder screening results among elementary school children in Japan. The test and screening results of 1033 children from a single elementary school between 2016 and 2018 were examined. Multiple regression analysis was performed for each gender of children of grades third and fourth to generate receiver-operating characteristic curves. Of the 69 children with parent-identified locomotor problems, 29 (42%) had physical fitness test score of mean ± 2 SD. For the standing long jump test in fourth grade girls, the results differed significantly (P < .001; cut-off, 127 cm) for children whose parents did (n = 7) and did not (n = 84) suspect a possible locomotive disorder. Physical fitness testing in combination with school-based screening for locomotive disorders may be useful for identifying and accurately diagnosing these disorders in children.