Root perforation is an adverse event that may accidentally occur during root canal treatment and can adversely affect the treatment plan and tooth prognosis. This study aimed to assess the accuracy of cone beam computed tomography (CBCT) for detecting the strip and furcal perforations with different sizes. The mesiolingual canals of 155 extracted human mandibular first molars were instrumented and randomly divided into four experimental groups and one control group (n = 31). Furcal (in the pulp chamber floor) and strip perforations with 0.5, 1, 1.5, and 2mm in diameter were manually created. The teeth were randomly mounted in bovine ribs and scanned using CBCT. Two radiologists unaware of the study groups observed the images and reported the greatest perforation diameter. The inter-observer agreements were calculated. The diagnostic accuracy was compared for furcal and strip perforations with different sizes. The inter-observer agreement for detecting furcal and strip perforations with different sizes were good. The diagnostic accuracy was 100% for the absence of perforation in each group. CBCT can detect the absence of perforation with high accuracy. The diagnostic accuracy of CBCT in the presence of strip and furcal perforation with different sizes was not significant.