Considering the rise of online education and an increasing number of students with disabilities in higher education, examining the validity of the Self-efficacy Questionnaire for Online Learning (SeQoL) for students with disabilities is warranted. The purpose of this study is to examine the reliability and validity of (SeQoL; Shen et al., 2013) for students with disabilities in higher education. We analyzed the internal structure, convergent validity, criterion validity, and reliability of SeQoL. A sample of 278 students with disabilities responded to an online survey in Spring 2021. Most of our sample were female, White, and undergraduate students. We used confirmatory factor analysis, correlation, multivariate analysis of variance, and Cronbach's alpha to analyze the data. Our results indicated that data fit the five factors model with 25 items. Students who preferred online or hybrid courses had significantly higher online learning self-efficacy than face-to-face courses. Limitations and future research were discussed.