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The accuracy of melanoma-specific dermoscopic criteria has been tested mainly in studies including invasive tumors. Scarce evidence exists on the usefulness of these criteria for the diagnosis of melanoma in situ (MIS).To investigate the diagnostic accuracy of dermoscopic criteria for the diagnosis of MIS.A diagnostic accuracy study with retrospective patient enrollment was conducted in 3 centers specializing in skin cancer diagnosis and management. A total of 1285 individuals with histopathologically diagnosed MIS or other flat, pigmented skin tumors that were histopathologically diagnosed or monitored for at least 1 year were included. Dermoscopic images of MIS and other flat, pigmented skin tumors were evaluated by 3 independent investigators for the presence of predefined criteria. Evaluators were blinded to the clinic dermoscopic and histopathologic diagnosis.Frequencies of dermoscopic criteria per diagnosis were calculated. Crude odds ratios, adjusted odds ratios, and corresponding 95% CIs were calculated by univariate and multivariate logistic regression, respectively.A total of 1285 patients were included in the study (642 [50%] male); mean age was 45.9 years (range, 9-91 years). Of a total of 1285 lesions obtained from these patients, 325 (25.3%) were MIS; 574 (44.7%) were nevi (312 [24.3%] excised and 262 [20.4%] not excised); 67 (5.2%) were seborrheic keratoses, solar lentigines, or lichen planus–like keratoses; 91 (7.1%) were pigmented superficial basal cell carcinomas; 26 (2.0%) were pigmented intraepithelial carcinomas; 100 (7.8%) were Reed nevi; and 102 (7.9%) were invasive melanomas with a Breslow thickness less than 0.75 mm. The most frequent dermoscopic criteria for MIS were regression (302 [92.9%]), atypical network (278 [85.5%]), and irregular dots and/or globules (163 [50.2%]). The multivariate analysis revealed 5 main positive dermoscopic indicators of MIS: atypical network (3.7-fold; 95% CI, 2.5-5.4), regression (4.7-fold; 95% CI, 2.8-8.1), irregular hyperpigmented areas (5.4-fold; 95% CI, 3.7-8.0), prominent skin markings (3.4-fold; 95% CI, 1.9-6.1), and angulated lines (2.2-fold; 95% CI, 1.2-4.1). When compared only with excised nevi, 2 of these criteria remained potent MIS indicators, namely, irregular hyperpigmented areas (4.3-fold; 95% CI, 2.7-6.8) and prominent skin markings (2.7-fold; 95% CI, 1.3-5.7).Clinicians should take into consideration the aforementioned dermoscopic indicators for the diagnosis of MIS.