Prostate puncture biopsy is the main clinical tool for the diagnosis of prostate cancer. The method of prostate puncture has undergone many changes, and in 1989, H odge et al. first reported a transrectal ultrasound-guided prostate puncture biopsy (6-stitch method), which has since been widely used in clinical practice. The detection rate of prostate cancer in patients with PSA between 4 and 10 ng/mL was reported to be 20%-32% by the 6-stitch transrectal prostate puncture biopsy, and the detection rate of prostate cancer increased with the increase of the number of puncture needles. The detection rate of prostate cancer increases as the number of puncture needles increases, while the method of trans-perineal prostate puncture biopsy is less used because of its tedious operation. The incidence of prostate cancer increases with the increase of PSA density, and some authors have reported that prostate puncture biopsy should be performed when the prostate PSA density is greater than 0.15 or 0.20 to exclude the possibility of prostate cancer. The principle is to represent the whole organ of the prostate with as few puncture sites as possible. We selected patients with serum tPSA between 4 and 20 ng/mL and comparable PSA densities and used ultrasound-guided prostate puncture biopsies at 12 sites via the rectal route and 14 sites via the perineal route, respectively. In addition, the use of a prostate-specific puncture template improved the accuracy of prostate tissue localization. The results showed that the positive rate of prostate puncture biopsy was approximately the same in both groups, indicating that trans-perineal prostate puncture biopsy did not affect the detection rate of prostate cancer compared with the conventional 12-point transrectal ultrasound-guided prostate puncture method. The positive puncture rate was higher in the transperineal group than in the transrectal group, especially for the prostatic metastases, which was mainly due to the fact that 4 stitches of the transperineal protocol were biopsies of prostatic metastases. The perineal prostate puncture biopsy method is better than the transrectal method for the detection of the peripheral zone of the prostate, where prostate cancer is more frequent. The tumors located in the migratory zone and the peripheral zone of the anterior prostate are easily missed. In our study, the incidence of tumors in the migratory zone was found to be 18%-24%, and the incidence of tumors completely in the migratory zone was found to be 2%-10%. Therefore, in our puncture protocol, biopsy of the metastatic zone of the prostate was added accordingly. In addition, because transrectal ultrasound-guided prostate puncture is performed from the posterior to the anterior part of the prostate, the biopsy of the anterior peripheral zone is easily missed in larger prostates. The trans-perineal puncture method basically avoids the above-mentioned drawbacks, not only for the peripheral zone tissues of each area, but also for the prostatic migratory zone tissues with 4 separate needles, which reduces the possibility of missing the diagnosis. In addition, transrectal puncture may introduce pathogenic bacteria from the rectum into the prostate tissue, thus increasing the risk of acute prostatitis/urinary tract infection. The main complications associated with this technique include hematuria, rectal bleeding, infection and dyspareunia, with an incidence of 47.34% reported by Bian Wei et al. in China. In the present study, the incidence of urinary tract infection was 19.6% in the TR group and 1.2% in the TP group, so the trans-perineal approach was superior to the trans-rectal approach in terms of infectious complications. In conclusion, the detection rate of prostate cancer by trans-perineal ultrasound-guided prostate aspiration biopsy was approximately the same as that of trans-rectal ultrasound-guided prostate aspiration biopsy, and the post-penetration complications of the former method were significantly less than those of the latter.