This three-phase study aimed to modify the Concerns about Inclusive Education Scale (CIES) under the Physical Education (PE) setting and to determine if the modified scale was reliable and valid. A total of 645 pre-service teachers (522 males and 123 females) from three universities participated in this study. In Phase 1, the CIES was revised, translated, and renamed to CIES-PE. In Phase 2, the content validity of the CIES-PE was tested by six experts. In phase 3, the construct validity of the CIES-PE was established through exploratory and confirmatory factor analyses and group invariance test across samples with and without adapted PE training experience. Results supported a five-factor, 24-item CIES-PE scale. In conclusion, the CIES-PE may be utilised as a reliable and valid instrument to assess the concerns of pre-service PE teachers about teaching students with disabilities in general PE classes.