Abstract. The objective of the present study was to explore the clinical value and safety of trans-rectal ultrasound (TRUS)-guided transperineal prostate 12+X core biopsy in the diagnosis of prostate cancer. Patients who received a TRUS-guided transperineal prostate biopsy for suspected prostate cancer at the General Hospital of The People's Liberation Army between September 2009 and May 2014 were retrospectively analyzed, this consisted of 1,300 patients. These patients were randomly divided into the 12+X core group or the standard 12-core group. The mean age of all the patients was 70.5 years old. Levels of prostate-specific antigen, digital rectal examination, transrectal ultrasound and magnetic resonance imaging (MRI) were checked and used as reference prior and subsequent to the biopsy procedure. The 12+X core group consisted of 937 patients and the 12-core group consisted of 363 patients. The mean number of core samples taken from both groups was 14.5 (ranging from 12 to 24) and the mean operative time of the whole group was 20.4 min (ranging from 15 to 40 min). The puncture positive detection rate of abnormal rectal examination, trans-rectal ultrasound, and MRI was 24.0, 30.1, and 59.2%, respectively, whereas the puncture positive rate was 47.2% in 12+X core group and 34.5% in 12-core group. Improved prostate needle biopsy with 12+X cores was found to have significantly higher detection rate than that with 12 cores as well as fewer post-operative complications, therefore making the method ideal for diagnosing prostate cancer.