INTRODUCTION: Currently, there are no established guidelines for follow-up (FU) after negative prostate biopsies (PBx) despite the presence of a target on MRI. We aimed to evaluate the risk of clinically significant prostate cancer (csPCa) within 2 years (2y) and at final FU in such cases. PATIENTS AND METHODS: We analyzed 105 patients with negative systematic and targeted PBx despite positive mpMRI (PI-RADS ≥ 3) (median FU: 66.5 months [IQR: 40-83]). All patients underwent FU with serial PSA measurements, digital rectal examinations and, when indicated, FU-MRI and/or FU-PBx. Outcomes assessed csPCa occurrence (GGG /ISUP ≥ 2) at 2y and at final FU, and predictive factors for csPCa. RESULTS: At 2y, the csPCa detection rate (Det-R) was 7.6%, increasing to 15.2% at final FU. No significant differences were observed at 2y based on baseline PI-RADS status. The mean initial PSAD was significantly higher in patients with csPCa at 2y versus without: 0.20 ng/mL² (SD: 0.11) versus 0.13 ng/mL² (SD: 0.12) (P = .008). Patients with baseline PSAD >
0.15 had a significantly higher 2y csPCa Det-R versus with PSAD <
0.15: 18.2% (4/22) versus 3.7% (3/81) (P = .036). The combination of PSAD <
0.15 and PI-RADS 3 at baseline was associated with a very low 2y csPCa Det-R (3%, 1/34), compared to PSAD >
0.15 and PI-RADS ≥ 4 (23%, 3/13). At final follow-up: 53% of patients with csPCa had an increasing PSAD (vs. 14% without, P = .003). A total of 41% (43/105) of patients underwent FU-MRI. Patients with csPCa were significantly more likely to have upgraded MRI findings (56% vs. 2.2%, P <
.002). CONCLUSION: Following negative PBx despite a positive mpMRI, the risk of csPCa was low. However, careful monitoring is essential, particularly in cases of PSAD >
0.15, and/or PI-RADS ≥ 4 at baseline. FU-PSAD and FU-MRI emerged as the most significant predictive factors, aiding to stratify the need for FU-PBx.