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The aim of this study was to check the in vitro accuracy of ICDAS criteria on digital images compared to visual examination for the diagnosis of occlusal caries against a micro-CT gold standard. ICDAS was scored in 40 extracted permanent molars by means of visual inspection and stereomicroscopic images. Visual examinations were performed in duplicate and at a one-week interval by three different calibrated examiners. The analysis of digital images by ICDAS criteria was also performed in duplicate, 1 month after visual examinations. The detection methods were compared by means of sensitivity, specificity, area under the curve, predictive positive and negative values, and accuracy for two different thresholds (1- sound vs. carious teeth; 2- tooth requiring operative vs. non-operative treatment). Sensitivity and accuracy values for threshold 1 in the visual ICDAS and image-based ICDAS methods were high for sensitivity (0.93 and 0.97) and for accuracy (0.83 and 0.85), but low for specificity (0.55 for both methods). Specificity values for threshold 2 were 0.77 and 0.82, while sensitivity was 0.33 and 0.28 for each method. Spearman's rank correlation coefficient was 0.53 and 0.43 (p<0.05) for visual and image-based ICDAS compared to the gold standard scores. Both visual and image-based ICDAS scores were similar to each other in terms of diagnostic accuracy when compared to the micro-CT gold standard. Low specificity for the presence of caries and sensitivity for the detection of caries requiring operative treatment were found. [ABSTRACT FROM AUTHOR]