The invention discloses a high-accuracy surface plasmon resonance (SPR) detection method. In the method, signal acquisition is performed on the same SPR detection system for multiple times under the same experimental condition to acquire a plurality of SPR curves with micro random translation, relative micro translation among the plurality of SPR curves is estimated, and an expected absolute position vector is further estimated on the basis, so that position uncertainty in the acquisition process of the SPR curves is avoided; meanwhile, by fusing different information among the plurality of SPR curves with the micro translation, a more accurate SPR curve with higher sampling density is acquired through estimation at the expected absolute position, so that the accuracy of the acquired SPR sensing information and the detection limit of an SPR system are improved.