Prostate-specific membrane antigen (PSMA)-targeted PET imaging recently emerged as a new method for the staging and restaging of prostate cancer. Most published studies investigated the diagnostic potential of Ga-labeled PSMA agents that are excreted renally. F-PSMA-1007 is a novel PSMA ligand that has excellent preclinical characteristics and that is only minimally excreted by the urinary tract, a potential advantage for pelvic imaging. The aim of this study was to investigate the diagnostic efficacy of F-PSMA-1007 for biochemical recurrence (BCR) after radical prostatectomy. From 3 academic centers, 251 patients with BCR after radical prostatectomy were evaluated in a retrospective analysis. Patients who had received second-line androgen deprivation therapy (ADT) or chemotherapy were excluded, but prior first-line ADT exposure was allowed. The median prostate-specific antigen (PSA) level was 1.2 ng/mL (range, 0.2-228 ng/mL). All patients underwent PSMA PET/CT at 92 ± 26 min after injection of 301 ± 46 MBq of F-PSMA-1007. The rate of detection of presumed recurrence sites was correlated with the PSA level and original primary Gleason score. A comparison to a subset of patients treated previously with ADT was undertaken. Of the 251 patients, 204 (81.3%) had evidence of recurrence on F-PSMA-1007 PET/CT. The detection rates were 94.0% (79/84), 90.9% (50/55), 74.5% (35/47), and 61.5% (40/65) for PSA levels of greater than or equal to 2, 1 to less than 2, 0.5 to less than 1, and 0.2 to less than 0.5 ng/mL, respectively. F-PSMA-1007 PET/CT revealed local recurrence in 24.7% of patients ( = 62). Lymph node metastases were present in the pelvis in 40.6% of patients ( = 102), in the retroperitoneum in 19.5% of patients ( = 49), and in supradiaphragmatic locations in 12.0% of patients ( = 30). Bone and visceral metastases were detected in 40.2% of patients ( = 101) and in 3.6% of patients ( = 9), respectively. In tumors with higher Gleason scores (<=7 vs. >=8), detection efficacy trended higher (76.3% vs. 86.7%) but was not statistically significant ( = 0.32). However, detection efficacy was higher in patients who had received ADT (91.7% vs. 78.0%) within 6 mo before imaging ( = 0.0179). F-PSMA-1007 PET/CT offers high detection rates for BCR after radical prostatectomy that are comparable to or better than those published for Ga-labeled PSMA ligands.