Objective·To evaluate diagnostic value and advantages of 99mTc-MIBI single photon emission computed tomography integrated with X-ray computerized tomography (SPECT/CT) in diagnosing patients with primary hyperparathyroidism (PHPT). Methods·Clinical data, postoperative pathological diagnosis, and clinical follow-up results of 249 patients with PHPT who underwent 99mTc MIBI SPECT/CT for diagnosis were systematically reviewed and compared with neck ultrasound, blood index tests (according to the diagnostic criteria of WHOs that must meet both high PTH and hypercalcemia), and double phase planar imaging (intravenous injection of 99mTc MIBI 740 MBq and undergoing 30 min and 120 min of cervical and thoracic early and delayed double phase static plane acquisition) results. Results·For 249 patients, diagnostic sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of 99mTc-MIBI SPECT/CT for the diagnosis of PHPT were 89.9%, 88.1%, 90.4%, 85.0%, and 88.0%. Those of neck ultrasound were 82.7%, 57.8%, 74.3%, 69.3%, and 72.6%.Those of blood indexes were 91.4%, 82.6%, 87.1%, 80.4%, and 87.6%. Those of planar imaging were 77.9%, 89.0%, 90.1%, 75.8%, and 82.7%. Conclusion·99mTc-MIBI SPECT/CT has greater clinical diagnostic value of PHPT than neck ultrasound and 99mTc-MIBI double phase planar imaging and can accurately locate lesions, so as to facilitate the parathyroidectomy.