The calibration path of system-level calibration directly affects the incentive effect of the error term and thus the calibration accuracy. Currently, the planning of system-level calibration paths is predominantly designed based on personal experience, resulting in insufficient incentive for error terms, low calibration accuracy, and long calibration times. Therefore, this study proposes a system-level calibration optimal path planning method based on an improved Dijkstra's algorithm. First, the system-level calibration optimal path planning problem was modeled as a multi-fork regular root tree model, and the adaptability of Dijkstra's algorithm was improved. Second, a 30-dimensional Kalman filter model was designed for system-level calibration. Then, simulation experiments were conducted, and the results demonstrated that the calibration accuracy of the error term reached 90% within 330 s. Finally, a Micro-Electro-Mechanical system (MEMS) inertial sensor, model PA-IMU488B, was used for experimental verification, and the results were compared with the discrete calibration results. The results indicate that the bias and scale factor errors of the MEMS inertial sensor reached the target accuracy within 5 min. The optimal path planning method for system-level calibration proposed in this study is not dependent on a high-precision turntable, is applicable to sensors of different accuracies, and decreases calibration time while ensuring calibration accuracy.