Objective To explore the value of combined application of PTH measurement and 99Tcm-sestamibi imaging in diagnosis of chronic renal failure. Methods 83 patients of chronic renal failure in our hospital were selected for measurement serum levels of PTH, Ca and P and for parathyroid imaging. Results The serum levels of PTH in patients in chronic renal failure were obviously higher than that of healthy controls (P<0.01). There were significant difference on the serum levels of Ca and P between patients with chronic renal failure and healthy controls (P<0.05). In dual-phase 99Tcm–MIBI parathyroid imaging for 83 patients with chronic renal failure, 72 lesions of parathyroid hyperplasia were found in 69 patients (3 patients had 2 hyperplasia lesions and 66 patients had 1 hyperplasia lesion). Conclusion The detection of serum PTH in patients with chronic renal failure could help in understanding patient’s condition and providing corresponding treatment, which is also significant in evaluation of curative effect and prognosis. The parathyroid imaging before surgery can be adopted to identify diagnosis and providing guidance for surgery treatment through location to lesion.