Objective High Resolution Computed Tomography (HRCT) has commonly been used for assessing the suspected cholesteatoma of temporal bone. Recently, Magnetic Resonance Imaging (MRI) has showed exciting performance on differentiating cholesteatoma from non-specific soft tissue. The aim of this study was to compare the accuracy of HRCT and MRI in the qualitative diagnosis of temporal bone cholesteatoma.Methods Patients with clinically suspected cholesteatoma were recruited in this study from September 2017 to March 2019 in the hospital. The participants were 107 cases (125 ears) aged from 3 to 79 years with an average age of 40.14 ± 18.8 years, including 67 males and 40 female. HRCT and MRI scans of temporal bone were routinely performed 3 days to 2 weeks before operation. The imaging scans were assessed by a radiologist and otological surgeon independently and the results were marked as suspicious cholesteatoma and noncholesteatoma. Intraoperative findings were recorded as cholesteatoma and noncholesteatomatous inflammatory tissues, which were confirmed by histopathology. Results The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) of HRCT was 68.8%, 39.6%, 64.6% and 44.2%, respectively, while for MRI was 96.1%, 91.7%, 94.9% and 93.6% respectively. The accuracy of MRI-DWI in the diagnosis of recurrent cholesteatoma was significantly higher than HRCT (P < 0.05). Conclusion The accuracy of MRI-DWI in the diagnosis of cholesteatoma is significantly higher than HRCT, especially in recurrent cases.