Objective To analyze the gray scale ultrasonography features and contrast enhanced imaging manifestations of 120 patients with ampulla lesions by double contrast enhanced ultrasound (DCEUS), and judge the diagnostic value of DCEUS for ampulla lesions. Methods Retrospective analysis of clinical data of 120 patients with ampulla benign and malignant lesions received DCEUS from January 2016 to June 2018 in our hospital was performed. The clinical data and characteristics of conventional grayscale ultrasound and contrast enhancement ultrasound imaging were analyzed. Results The pathological results of 120 cases showed that 79 cases were malignant lesions and 41 cases benign lesions. Among these malignant lesions, there were 20 cases of ampulla carcinoma, 26 cases of lower common bile duct carcinoma, 30 cases of pancreatic head carcinoma and 3 cases of duodenal carcinoma. Among these benign lesions, there were 10 cases of ampullary inflammatory stenosis, 3 cases of pancreatic head cystic adenoma and 28 cases of lower choledocholithiasis. The age of benign group was younger than that of the malignant group (P＜0.05). Gray scale ultrasonography showed that there were significant differences between the two groups in terms of lesion boundaries, morphology, and main pancreatic duct expansion (P＜0.05). The proportions of clear boundaries, regular morphological and main pancreatic duct expansion in benign and malignant group were 68.3%, 73.2%, 14.6% and 15.2%, 27.8%, 59.5%, respectively. The area under ROC curve (AUC), sensitivity, specificity, positive predictive value and negative predictive value of DCEUS in diagnosing ampulla lesions were 0.981, 96.2%, 100%, 100% and 93.2%, respectively. Conclusion DCEUS could clearly and sensitively display the characteristics of ampulla benign and malignant lesions and local microcirculation perfusion and has a high diagnostic value and differential diagnostic value for ampulla benign and malignant lesions.