Self-reference detection is necessary and important to a biosensor. The linear weak measurement system based on total internal reflection has attracted widespread attention due to its high stability, label-free detection, and easy integration. In this paper, we propose a differential detection method based on the linear total internal reflection weak measurement system. We introduce the half-wave plate (HWP) to convert the H light and the V light to each other, thereby obtaining the difference in phase change of the optical path before and after the HWP. Experiments show that the system can not only achieve differential detection, but also has high stability. The linear differential weak measurement system proposed in this paper not only provides a new differential measurement method for real-time biosensors, but also enriches the types of weak measurement sensors.