The invention relates to the technical field of the road bridge engineering detection, and discloses a highway steel bridge fatigue analysis method based on an actually measured stress spectrum. The technical problem that the strain detection data collected through the current actual measurement method has multiple idle clutter is solved. The method comprises the following steps: S1, acquiring data; S2, processing a strain data signal: S21 filtering; S22, peak value extracting; S23, small stress circular filtering; S24, base line drift processing; S3, performing rain-flow method stress circular counting, thereby obtaining stress spectrum through the rain-flow method counting; and S4, evaluating a fatigue problem by utilizing a fatigue life evaluation theory, and computing fatigue damage and residual life of each measurement point. According to above technical scheme, the fatigue strain monitoring data is acquired and collected through an actual measurement method, and then the filtering, the peak extraction, the small stress circular filtering and the baseline drifting processing and like operations are performed on the acquired fatigue strain monitoring data, and finally, the fatigue damage and the residual life of each measurement point are computed, thereby reaching an aim of improving the evaluation accuracy and reliability.