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To assess the efficacy and safety of transperineal template-guided prostate biopsy.From December 2003 to December 2013, a total of 3 007 patients (30–91 years old, mean age 69.1) who met the inclusion criteria underwent 11-region transrectal ultrasound-guided transperineal template prostate biopsy.The inclusion criteria included a prostate-specific antigen (PSA) level of 4.0 ng/mL or greater and abnormal prostate gland findings on digital rectal examination, ultrasound, CT or MRI. The median PSA level was 11.0 ng/mL (range 0.2–100 ng/mL).The prostate cancer detection rate and prostate biopsy adverse effects, as well as prostate cancer spatial distribution were analyzed.A mean of 19.3 cores (range 11–44) were obtained for each biopsy, and more cores were obtained in larger prostates than in smaller ones.One to four cores were collected from each region. Prostate cancer was detected in 1 067 of the 3 007 patients (35.5%). The prostate cancer detection rates in groups with PSA levels of 0–4.0 ng/mL, 4.1–10.0 ng/mL, 10.1–20.0 ng/mL, 20.1–50.0 ng/mL, and 50.1–100.0 ng/mL were 15.3% (27/176), 21.0% (248/1 179), 32.6% (318/975), 56.0% (232/414), and 92.0% (241/262), respectively.The mean positives for cancer in regions 1–10 and region 11 (the apical region) were 46.7% vs 52.0% (P = 0.014).Regarding adverse effects, 47.0% of the patients reported hematuria, 6.1% developed hemospermia, 1.9% required short-term catheterization after biopsy because of acute urinary retention, and 0.03% (one patient) developed urosepsis.Transrectal ultrasound-guided transperineal template prostate biopsy is safe and accurate.The current study suggests that prostate carcinoma foci are more frequently localized in the apical region.