PURPOSE: To evaluate differences between autorefraction measurements with and without cycloplegia among school-aged individuals and to explore factors associated with significant differences. DESIGN: Cross-sectional, retrospective study. PARTICIPANTS: Individuals between 3 and 22 years of age evaluated at the Illinois College of Optometry from September 2016 through June 2019 who underwent same-day noncycloplegic and cycloplegic autorefraction of the right eye. METHODS: Demographic information including age, sex, and race or ethnicity were collected during the eye examination. Autorefraction was performed before and after cycloplegia. Myopia, defined as at least -0.50 diopter (D) spherical equivalent (SE), hyperopia, defined as at least +0.50 D SE, and astigmatism of at least 1.00 D cylinder were determined using noncycloplegic and cycloplegic autorefractions. Factors associated with at least 1.00 D more myopic SE or at least 0.75 D cylindrical difference by noncycloplegic autorefraction were assessed using logistic regression models. MAIN OUTCOME MEASURES: Differences between noncycloplegic and cycloplegic autorefraction measurements. RESULTS: The mean age was 10.8 ± 4.0 years for the 11 119 individuals; 52.4% of participants were female. Noncycloplegic SE measured 0.65 ± 1.04 D more myopic than cycloplegic SE. After adjusting for demographic factors and refractive error, individuals with at least 1.00 D of more myopic SE refraction by noncycloplegic autorefraction (25.9%) were more likely to be younger than 5 years (odds ratio [OR], 1.45; 95% confidence interval [CI], 1.18-1.79) and 5 to younger than 10 years (OR, 1.32; 95% CI, 1.18-1.48) than those 10 to younger than 15 years. This difference of at least 1.00 D of more myopic SE was more likely to be observed in Hispanic people (OR, 1.23; 95% CI, 1.10-1.36) and those with hyperopia (OR range, 4.20-13.31). Individuals with 0.75 D or more of cylindrical difference (5.1%) between refractions were more likely to be younger than 5 years, to be male, and to have mild-moderate-high myopia or moderate-high hyperopia. CONCLUSIONS: Three quarters of school-aged individuals had < 1 D of myopic SE difference using noncycloplegic compared with cycloplegic autorefraction. Understanding measurement differences obtained for refractive error and associated factors may provide useful information for future studies or programs involving refraction in school-aged children.