Background: A meta-analysis was performed to evaluate the efficacy and safety of intrarectal local anesthestic (IRLA), periprostatic nerve block (PPNB), and the combined modalities in alleviating the pain during transrectal ultrasound (TRUS)-guided prostate biopsy. Materials and methods: A literature review was performed to identify all published randomized controlled trials (RCTs) about IRLA vs no anesthesia or placebo gel; PPNB vs no injection, periprostatic placebo injection, or IRLA; combined PPNB and IRLA vs PPNB alone; and combined PPNB and intraprostatic nerve block (IPNB) vs PPNB alone before TRUS-guided biopsy. Sources included MEDILINE, EMBASE, and Cochrane Library from 1980 to 2016. The main outcomes were biopsy pain score, probe manipulation pain score, and anesthetic infiltration pain score assessed by the visual pain scale. Results: A total of 26 articles involving 36 RCTs were used in this analysis: Although IRLA can lead to pain reduction, the result was not statistically significant when compared with no anesthesia or placebo gel (weighted mean difference [WMD]: -0.22, 95% CI: -0.45 to 0, P=0.06). PPNB can lead to significantly lower biopsy pain scores when compared with no analgesia (WMD: -1.32, 95% CI: -1.68 to -0.95, P<0.00001), placebo injection (WMD: -2.62, 95% CI: -3.16 to -2.07, P<0.00001), or IRLA (WMD: -1.31, 95% CI: -1.40 to -1.22, P<0.00001). PPNB + IRLA can lead to significantly lower biopsy pain scores when compared with PPNB alone (WMD: -0.45, 95% CI: -0.62 to -0.28, P<0.00001). PPNB + IPNB can lead to significantly lower biopsy pain scores when compared with PPNB alone (WMD: -0.73, 95% CI: -0.92 to -0.55, P<0.00001). There were no severe reported general or local complications related to local anesthesia. Conclusion: This meta-analysis indicates that a combination of PPNB and IRLA/IPNB is effective and safe in alleviating the pain during TRUS-guided prostate biopsy. Further high-quality RCTs are needed to validate this result.