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Any of several conditions can cause asthenopia. The purpose of this study was to determine whether the particular sensations or their location vary dependent on the symptom-inducing condition.Twenty subjects with good vision performed eight reading tasks in random order during different conditions. Each condition used different stimuli to induce asthenopia. The eight conditions were mixed astigmatism, close viewing distance, upward gaze, dry eyes, lens flipper, small font, glare, and flickering light. Subjects were asked to read until attaining a level of discomfort self-defined as “barely tolerable.” After each task, subjects rated the magnitude of several symptom descriptors (burning, ache, strain, irritation, tearing, blurred vision, double vision, dryness, and headache) and their location.Analysis of variance with repeated measures was used to determine that all of the individual symptom sensation measures (except blur) were significantly related (p values ranged from 0.003 to <0.0001) to the inducing condition. Principal factor analysis with orthogonal varimax rotation was used to test symptom by condition relationships and determined two latent factors, designated external and internal symptom factors (ESF and ISF), that related symptoms to inducing condition. The ESF pattern comprises burning, irritation, tearing, and dryness located in the front and bottom of the eye. ESF is caused by holding the eyelid open, glare, up gaze, small font, and flickering. ESF seems highly related to dry-eye symptoms. The ISF pattern comprises ache, strain, and headache located behind the eyes. ISF is caused by the close viewing distance, lens flipper, and mixed astigmatism conditions and is likely related to accommodative and vergence stress.Symptom descriptors and locations were able to distinguish discomfort on the basis of causative condition. Results support two different symptom constellations and, hence, at least two different afferent pathways for symptoms of asthenopia.