Objective To assess the clinical value of SPECT/CT bone scan for differential diagnosis of spinal single positive lesions. Methods A total of 921 patients with bone diseases (412 males, 509 females，age 68.82±12.07 y) were included in this study. 99Tcm-MDP whole-body planar bone scan and regional SPECT/CT tomography were conducted. Results Among the 921 patients, 161 cases were diagnosed as metastatic tumor of bone (17.5%), 4 cases were diagnosed as primary bone tumors (0.4%), 2 cases were diagnosed as myeloma (0.2%), 5 cases were diagnosed as bone tuberculosis (0.5%), 689 cases were diagnosed as benign lesions (74.8%), 59 cases were diagnosed as traumatic osteoporotic vertebral compression fractures (6.4%), 1 cases was diagnosed as Schmorl's nodes (0.1%). A total of 161 patients were found with metastasis tumor of bone, including 9 cases (5.6%) of cervical, thoracic in 46 cases (28.6%), 82 cases (50.9%) of lumbar and sacral vertebra in 24 cases (14.9%). According to site classification, a total of 30 cases of cervical lesions, 9 cases with bone metastases (30%); a total of 224 cases of thoracic vertebral lesions, 46 cases with bone metastases (20.5%); lumbar lesions were 606 cases, 82 cases with bone metastases (13.5%); sacral vertebral lesions in 61 cases, 24 cases with bone metastases (39.3%). Conclusion SPECT/CT tomography provides accurate positioning of single positive lesions detected in the whole-body bone scan, and also shows the morphology and the imaging features of soft tissue around this lesion. It might significantly reduce the false positive rate of vertebral bone scan in single lesion, and has important clinical value in the differential diagnosis of single spinal lesions.