OBJECTIVE The aim of this study was to evaluate the role of three-dimensional transrectal ultrasound in the diagnosis of prostate cancer. METHODS A total of 112 patients with elevated serum prostate-specific antigen (PSA) or a positive digital rectal examination were evaluated using three-dimensional greyscale transrectal ultrasound (3D-GS TRUS) and three-dimensional power Doppler sonography (3D-PDS). Target biopsies were obtained together with 12 core systematic biopsies. Pathological results were correlated with the imaging data. RESULTS Cancers were detected in 269 biopsy sites from 41 patients. 229 sites of cancer were depicted by 3D-GS TRUS and 213 sites were depicted by 3D-PDS. 30 sites were missed by both 3D-GS TRUS and 3D-PDS. Abnormal prostate images depicted by 3D-GS TRUS and 3D-PDS were associated with lesions with a Gleason score of 6.9 or higher. CONCLUSIONS The detection rates of prostate cancer were significantly improved with 3D-GS TRUS and 3D-PDS on serum PSA levels >10 ng ml(-1) or 20 ng ml(-1). 3D-GS TRUS and 3D-PDS may improve the biopsy yield by determining appropriate sites for target and systematic biopsies. The abnormalities detected by 3D ultrasound were associated with moderate- and high-grade prostate cancers. However, based on the number of false-negative TRUS results, the use of systematic prostate biopsies should not be eliminated.