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Background Visual acuity is a critical component of visual function assessment for all ages. Standardized vision testing protocols may help prevent testing inconsistencies resulting from variations in test administration and interpretation of different examiners. However, most vision assessments outside of research settings, including in doctor’s offices, rarely employ standardized protocols. Validated protocols such as the Early Treatment for Diabetic Retinopathy Study (ETDRS) are frequently employed by vision researchers to ensure accurate and repeatable visual acuity measurements. Methods This study evaluates a desktop-based standardized vision testing algorithm (EyeSpy 20/20) specifically designed for use on mobile electronic platforms. Subjects were tested on a desktop version of the EyeSpy software for both the accuracy and duration of measurement of visual acuity and compared to a e-ETDRS chart in a randomized sequence. Children were recruited for this study and tested between two different visual acuity measurement systems. Bland–Altman analysis and correlation tests were done. Results Hundred and ten children were recruited for the study. The EyeSpy 20/20 visual acuity testing algorithm as tested with the desktop version was non-inferior to the gold standard e-ETDRS testing algorithm on a desktop platform, but statistically faster to implement when administered on the same electronic testing platform. Conclusion EyeSpy 20/20 is a promising tool for vision screening and visual acuity evaluation in children.

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