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Engineering applications have shown that variable-section supports of steel crane girders are prone to fatigue fracture, and traditional fatigue prediction experience is not applicable to these types of supports due to the complex internal stresses caused by their unique geometries. In this paper, a new fatigue prediction approach was proposed for variable-section supports of steel crane girders based on the peak stress method; this approach can locate the fatigue initiation sites and predict the fatigue lives of variable-section supports under axial and bending loading. Then, some constant-amplitude fatigue test results were presented for typical variable-section supports of steel crane girders. A comparison was made between the experimental results and theoretical estimations using the peak stress method. The results indicated that the peak stress method is appropriate to estimate fatigue life up to crack initiation in variable-section supports of steel crane girders subjected to axial and bending loads.