To compare the diagnostic accuracy of [F]-Fluorocholine (FCH) PET/CT with conventional [mTc]Tc-MIBI scintigraphy and cervical ultrasound (USG) for the preoperative localization of hyperfunctioning parathyroid tissue (HFPT) in patients with primary hyperparathyroidism (PHPT). This prospective study included 90 patients diagnosed with PHPT who underwent [F]F-CH PET/CT, [mTc]Tc-MIBI SPECT/CT and Neck USG. The diagnostic accuracy of each imaging modality was assessed using intraoperative findings and histopathological confirmation as the gold standard.