To compare the role of 99mTc-MIBI SPECT-CT image and US in reoperation of persistent secondary hyperparathyroidism patients. A total of 8 persistent secondary hyperparathyroidism patients underwent parathyroidectomy. The sensitivity and accuracy of US and 99mTc-MIBI images before operation were determined. 9 of 14 surgical resection tissues from 8 persistent secondary hyperparathyroidism patients were confirmed to parathyroid hyperplasia. The results showed that the sensitivities were 77.8% and 100%, respectively, for US and 99mTc-MIBI SPECT-CT images. And the accuracies of US and 99mTc-MIBI dual time planar image and SPECT-CT were 50%, 78.6%, respectively. There was significantly difference between two procedures (P=0.021). The superior and inferior localization of glands were both detected by the ultrasound and scintigraphy. Two ectopic parathyroidism nodules were found by 99mTc- MIBI SPECT-CT imaging, while US provided no consistent findings. There was no significantly difference between two procedures (P=0.3). Although two cases of them were not confirmed by pathology, the iPTH of them (800 and 1429 respectively) were much higher than other four cases (<400) pg/ml. PTX is a safe and effective treatment for the patients with persistent SHPT. 99mTc-MIBI planar and SPECT-CT imaging may provide more helps for clinician’s localization the hyperparathyroidism glands accurately.