Objective: To explore the feasibility and application value of the reduction of the number of needle punctures in the positive area of contrast-enhanced ultrasonography (CEUS) in prostate needle biopsy. Methods: We selected 120 patients who had positive areas with suspected prostate cancer (PCa) in CEUS in the Affiliated Hangzhou First People’s Hospital, Zhejiang University School of Medicine. Two different puncture methods were randomly used for biopsy, including 48 patients with 10-point systematic puncture, and the other 72 patients with reduced-needle puncture in contrast-positive area. All cases were confirmed by surgery and pathology. The detection rate and complications of the 2 methods were compared and analyzed. Results: Among the 120 patients, 76 were diagnosed with PCa (76/120, 63.3%), the detection rate in the systemic puncture group was 64.6% (31/48), and the detection rate in the reduced-needle puncture group was 62.5% (45/72). There was no statistically significant difference in the detection rate of PCa between the 2 groups (P=0.817). A total of 311 tissues were obtained from 31 PCa patients in the systematic puncture group, of which 126 (40.5%) were positive, and a total of 372 tissues were obtained from 45 PCa in the reduced-needle puncture group, of which 276 (74.2%) were positive, with a statistically significant difference in the number of positive needle punctures between the 2 groups (P<0.001). The incidence of complications such as hematuria in the systemic puncture group was higher than that in the reduced-needle puncture group (20.8% vs 6.9%, P=0.024). Conclusion: When a large positive area was detected in prostate by CEUS, the use of appropriate reduced-needle puncture does not significantly reduce the detection rate of prostate cancer, but improve the rate of positive needle punctures and reduce the complications induced by puncture, which has certain clinical application value.