Objective: To evaluate the effects of three different local anesthetic agents in patients who underwent Transrectal ultrasoun guided prostate needle biopsy. Materials and Methods: One hundred and sixty patients who were admitted to our clinic between January 2012 and May 2012 for prostate biopsy were divided into 4 groups: no anesthesia-administered group (group 1), lidocaine-administered group (group 2), levobupivacaine-administered group (group 3) and bupivacaine-administered group (group 4). Pain intensity was evaluated in each group using the visual analog scale (VAS): during administration of local anesthetic (VAS 1), during the biopsy (VAS 2), just after the biopsy (VAS 3) and one hour after the biopsy (VAS 4). Results: There was not any significant difference among the groups in terms of VAS 1 (p=0.152). Pain scores were significantly lower during biopsy (VAS 2) in groups 1, 2 and 3 compared to that in group 1 (p=0.001, p=0.005 and p=0.007, respectively). VAS 3 scores were significantly lower in group 2 and group 3 compared to group 1 while no difference was found in group 4 (p=0.003, p=0.032, and p=0.136, respectively). VAS 4 scores were significantly lower only in group 3 compared to group 1 (p=0.001). Conclusions: It was observed that all the three local anesthetics effectively diminished pain during prostate biopsy. However, we found that levobupivacaine, which improved pain scores in all steps of pain evaluation compared to the control group, was relatively superior for pain management in the transrectal biopsy setting.