Objective: To compare of positive results and puncture complication rate of transperineal and transrectal prostate puncture in different prostate-specific antigen (PSA) intervals by biplane B-ultrasound guided. Methods: Retrospectively analyzed the clinical data of 629 patients who underwent double-plane B-ultrasound guided prostate puncture from January 2017 to December 2018 at the First Affiliated Hospital of University of Science and Technology of China, including 282 patients in the perineal group and 347 patients in the rectal group. The PSA value of the patients was divided into ≤ 4 ng/mL, 4 ng/mL < PSA ≤ 10 ng/mL, 10 ng/mL < PSA ≤ 20 ng/mL, 20 ng/mL < PSA < 100 ng/mL and ≥100 ng/mL 5 intervals, respectively and total puncture positive rate of the 5 intervals were counted. Basic data such as age, prostate volume, body mass index (BMI), Gleason score, and complications were counted, and differences between the two groups were compared. Results: The overall positive rate of the perineal group was 52.13% and of the rectal group was 40.63%, they had significant difference (P < 0.05). In the 5 PSA intervals, there were no significant difference about positive rates of the two groups (P > 0.05). But in the PSA 20~100 ng/mL interval, the positive rates have significant differences (P < 0.05). About the overall complications, the rectal group was higher than the perineal group, the difference was statistically significant (P < 0.05). Among them, gross hematuria, urinary tract infection, fever, and vasovagal reaction were significantly higher in the rectal group (P < 0.05), except acute urinary retention (P > 0.05). Rectal hemorrhage was a special complication of the rectal group, and the perineal swelling and pain were specific complications of the perineal group. Conclusion: The positive rate of prostate puncture in the perineal group is significantly higher than in the rectal group. The incidence of complications in the perineal group is significantly lower than in the rectal group.