OBJECTIVE: This study evaluated the accuracy in localisation and distribution of real-time three-dimensional (4-D) ultrasound-guided biopsies on a prostate phantom. METHODS: A prostate phantom was created. A three-dimensional real-time ultrasound system with a 5.9MHz probe was used, making it possible to see several reconstructed orthogonal viewing planes in real time. Fourteen operators performed biopsies first under 2-D then 4-D transurethral ultrasound (TRUS) guidance (336 biopsies). The biopsy path was modelled using segmentation in a 3-D ultrasonographic volume. Special software was used to visualise the biopsy paths in a reference prostate and assess the sampled area. A comparative study was performed to examine the accuracy of the entry points and target of the needle. Distribution was assessed by measuring the volume sampled and a redundancy ratio of the sampled prostate. RESULTS: A significant increase in accuracy in hitting the target zone was identified using 4-D ultrasonography as compared to 2-D. There was no increase in the sampled volume or improvement in the biopsy distribution with 4-D ultrasonography as compared to 2-D. CONCLUSION: The 4-D TRUS guidance appears to show, on a synthetic model, an improvement in location accuracy and in the ability to reproduce a protocol. The biopsy distribution does not seem improved.