Objective. To evaluate the pain score during transrectal ultrasound (TRUS)-guided prostate biopsy using three different anaesthetic applications and no anaesthesia. Material and methods. One-hundred men undergoing TRUS-guided prostate biopsy were prospectively enrolled in this study. Patients were randomized to four groups. Group 1 (n = 25) received no anaesthesia, group 2 (n = 25) was administered a perianal lidocaine injection, group 3 (n = 25) was administered a periprostatic lidocaine injection, and group 4 (n = 25) was administered a combination of perianal intrarectal lidocaine prilocaine cream. The anaesthetic application was given 5 min before the procedure. All patients were asked to indicate the level of pain experienced on a visual analogue scale (VAS) during three situations, including when the TRUS probe was inserted and 15 min and 2 weeks after biopsy. Results. Mean pain scores were similar in all groups at the time of probe insertion. Mean pain scores showed statistically significant differences between group 1 and the other groups, except for group 4, 15 min after the procedure. Group 3 showed better pain control 15 min after biopsy and this difference was statistically significant (p < 0.043). The VAS scores were similar 2 weeks after the procedure in all groups. Conclusions. Anaesthetic application before TRUS-guided prostate biopsy may be advocated. The application of periprostatic lidocaine seems to be the most advantageous method for lowering the perception of pain.