Ambulance ride comfort not only impacts health of patients, but influences handling capability of drivers. Previous studies on ambulance ride comfort mainly focused on analysis and optimization of ride comfort of supine patients, but ride comfort of drivers was ignored. For solving this problem, a 10°‐of‐freedom (DOF) vibration model of ambulance was established, and a supine patient on stretcher and a driver on seat were included in the model. The excitations on front and rear wheel were proposed according to speed bump, which have a great impact on occupants. The objective function and design variables of multi‐objective optimization were determined, and the parameters of design variables were solved by using Elitist Nondominated Sorting Genetic Algorithm (NSGA‐II). Based on the parameters before and after optimization, ride comfort of supine patient and driver was analyzed. The results show that ride comfort of supine patient and driver in ambulance under speed bump was improved 20% and 7%, respectively, and ride comfort of supine patient and driver in ambulance under random road was increased by 9% after optimization. Multi‐objective optimization of ambulance ride comfort under speed bump can improve ride comfort of supine patient and driver, and has a good optimal effect. © 2019 Institute of Electrical Engineers of Japan. Published by John Wiley & Sons, Inc.