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Study Type – Diagnostic (exploratory cohort)Level of Evidence 2bWhat's known on the subject? and What does the study add?Template assisted transperineal biopsy of the prostate has become increasingly popular over the past decade. Several studies have demonstrated that transperineal prostate biopsy (TPB) is associated with an increased rate of cancer detection, increased histological concordance with final prostatectomy samples and an increase in anterior and apical prostate cancers than standard TRUS biopsy. However, interpretation of the literature is difficult due to considerable variation between studies in terms of technique and equipment.We examined a small cohort (n= 40) of patients using a standardized 36 core template assisted TPB technique. We show that utilising this technique is associated with high cancer (68%) detection rate in patients with two previous negative TRUS biopsies. Of patients were found to have anterior gland tumours which would not have been detected by standard TRUS guided biopsy.To determine the efficacy and safety of a standardized 36 core template-assisted transperineal biopsy technique for detecting prostate cancer in patients with previously negative transrectal ultrasonography-guided prostate biopsies and elevated prostate-specific antigen (PSA) levels.Between April 2008 to September 2010, a total of 40 patients with a mean (range) age of 63 (49–73) years, a mean (range) elevated PSA level of 21.9 (4.7–87) ng/mL and two previous sets of negative TRUS-guided prostate biopsies underwent standardized 36 core template-assisted transperineal prostate biopsies under general anaesthetic as a day case procedure.The cancer detection rate and complications for all cases were evaluated.In total, 27 of 40 (68%) patients were found to have adenocarcinoma of the prostate, two patients (5.0%) had atypical small acinar proliferation, one had high-grade prostatic intraepithelial neoplasia (2.5%), four (10%) had chronic active inflammation and six (15%) had benign histology.Gleason scores were in the range 6–9, with a median Gleason score of 7.There were no cases of urosepsis, urinary tract infections or haematuria. A single patient experienced acute urinary retention, with a subsequent succesful trial without a catheter, and haematospermia was common, although minor.Our standardized 36 core template-assisted transperineal prostate biopsy technique is safe and associated with a high detection rate of prostate cancer.This technique should be considered in patients with elevated PSA levels and previously negative TRUS-guided prostate biopsies.