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Predominantly peripheral disease in eyes with nonproliferative diabetic retinopathy (DR) is suggested as a potential strong risk factor for progression to proliferative disease. However, the reliability and optimal method for the assessment of lesion distribution are still uncertain.To compare agreement between subjective assessment and precise quantification of lesion burden in ultrawidefield (UWF) images of eyes with DR.This multisite cross-sectional study examines UWF pseudocolor images acquired from DR screening clinic patients from December 20, 2014, through August 1, 2014. Of 104 cases, 161 eyes with DR were included. Data analysis was conducted from June 1, 2016, through December 1, 2016 at the Doheny Image Reading Center.Distribution of DR lesions in eyes was assessed subjectively and quantitatively, and eyes were classified as having predominantly central lesions (PCLs) or predominantly peripheral lesions (PPLs). The frequency and surface area (SA) of each lesion type were quantified. Intergrader and subjective vs quantitative classification were compared for level of agreement. Several methods of determining PPL distribution were also compared.On subjective frequency-based evaluation by graders, 133 eyes were classified as having PCL, and 28 eyes as having PPL. On exact quantification of lesion SA, 121 eyes were classified as PCL, and 40 eyes as having PPL. On SA-based quantification, 134 eyes were classified as having PCL, and 27 eyes as having PPL. There was a significant difference between qualitative and quantitative classification of DR lesion distribution for both frequency-based (mean difference [SD]: PCL, 6 ; PPL, 13 ; P < .001) and SA-based (mean difference [SD]: PCL, 6 ; PPL, 20 ; P < .001) methods. Both intergrader reproducibility and subjective vs quantitative agreement were higher with frequency-based classification.Subjective assessment of PPL DR lesions on UWF images differed in some cases from precise quantitative assessments, particularly when considering the area of lesions. These findings highlight the benefit of objective quantitative approaches to DR assessment, which may facilitate the development of a more precise DR scoring system.