The aim of this study was to review the efficacy of contrast-enhanced harmonic endoscopic ultrasonography (CH-EUS) in diagnosing gallbladder (GB)-protruded lesions. Thirty-eight patients underwent CH-EUS for the diagnosis of GB-protruded lesions. Twenty-four patients whose major axes of their largest lesions were longer than 10 mm were recruited. The ability of CH-EUS to diagnose malignant or benign lesions was reviewed. We treated lesions with brindled enhanced patterns as malignant and those with uniformly enhanced or unenhanced patterns as benign. Furthermore, three gastroenterologists who were not familiar with pancreaticobiliary EUS compared the diagnostic abilities of CH-EUS and conventional EUS using photographs. The sensitivity, specificity, and malignant accuracy of CH-EUS were 100, 94.4, and 95.8%, respectively. The number of lesions that presented with enhanced patterns was significantly different between the malignant lesions and the benign lesions (P < 0.001). In the comparison of diagnostic abilities between CH-EUS and conventional EUS by the three gastroenterologists, CH-EUS was significantly superior to conventional EUS in terms of sensitivity, specificity, and accuracy (76.1 vs. 42.9%, P = 0.029; 66.7 vs. 39.2%, P = 0.005; and 69.4 vs. 40.3%, P < 0.001; respectively). In conclusion, CH-EUS was useful for diagnosing malignant and benign GB-protruded lesions.