To assess the diagnostic accuracy of cone beam computed tomography (CBCT) in comparison with conventional radiography for vertical root fractures, 50 of 100 teeth were subjected to vertical root fracture (VRF) and then placed in dry mandibles. 3D scans were obtained for all teeth, and conventional radiographs were used as control images. All the images were assessed by 6 observers, who determined the presence of root fractures by using a 5-point confidence rating scale. The mean area under the curve (Az) for CBCT was 0.91, and that for conventional radiography was 0.64. The difference between the modalities was statistically significant (P = 0.003). On the basis of interclass coefficient, inter-observer agreement for CBCT was 0/750, and that for conventional radiography was 0/637. Thus CBCT was shown to be significantly better than conventional periapical radiography for diagnosis of vertical root fractures in vitro. (J Oral Sci 52, 593-597, 2010)