Shear wave elastography (SWE) is currently used to detect tissue pathologies and, in the setting of preventive medicine, may have the potential to reveal structural changes before they lead to functional impairment. Hence, it would be desirable to determine the sensitivity of SWE and to investigate how Achilles tendon stiffness is affected by anthropometric variables and sport-specific locomotion. To investigate the influence of anthropometric parameters on Achilles tendon stiffness using SWE and examine different types of sports to develop approaches in preventive medicine for professional athletes, standardized SWE of Achilles tendon stiffness was performed in 65 healthy professional athletes (33 female, 32 male) in the longitudinal plane and relaxed tendon position. Descriptive analysis and linear regression were performed. Furthermore, subgroup analysis was performed for different sports (soccer, handball, sprint, volleyball, hammer throw). In the total study population (n = 65), Achilles tendon stiffness was significantly higher in male professional athletes (p < 0.001) than in female professional athletes (10.98 m/s (10.15-11.65) vs. 12.19 m/s (11.25-14.74)). Multiple linear regression for AT stiffness did not reveal a significant impact of age or body mass index (BMI) (p > 0.05). Subgroup analysis for type of sport showed the highest AT stiffness values in sprinters (14.02 m/s (13.50-14.63)). There are significant gender differences in AT stiffness across different types of professional athletes. The highest AT stiffness values were found in sprinters, which needs to be considered when diagnosing tendon pathologies. Future studies are needed to investigate the benefit of pre- and post-season musculoskeletal SWE examinations of professional athletes and a possible benefit of rehabilitation or preventive medicine.