Objective: The aim of our research was to evaluate the usefulness of real-time ultrasound elastography (RTE) in the differential diagnosis between benign and malignant non-palpable and palpable testicular lesions in children and adults. Methods: B-mode ultrasonography, color/power Doppler, and RTE of 54 patients with 54 unilateral testicular lesions were retrospectively investigated. The imaging results were supported by histopathological examination in all cases. On RTE, the value of tissue rigidity was reported in a chromatic scale and categorized into three patterns: Soft (red), medium (green), and hard (blue). Results: Among the 54 testicular lesions, 35 measured more than 2 cm in diameter (32 malignant and 3 benignant tumors) while 19 were <2 cm (9 malignant and 10 benignant lesions). In the group of the 41 proved malignant tumors, RTE demonstrated hard pattern in 32 cases. Immature teratomas exhibited a mixed pattern, likely related to their heterogeneous content. In the testicular lesion diagnosis, RTE showed a 78% sensitivity, an 85% specificity, a 94% positive predictive value, a 55% negative predictive value, and an 80% accuracy in differentiating malignant from benignant lesions. The integration of ultrasound (US) with color/power Doppler and RTE showed a diagnostic accuracy of 100% and 90%, respectively. Conclusion: Integrated with conventional US methods, RTE increases the diagnostic accuracy of US and color/power Doppler in the differential diagnosis between benignant and malignant testicular lesions.