As an innovative cement-based material, the ultra-high performance concrete (UHPC) has been recently utilized to reinforce the orthotropic steel bridges. Based on an in-situ steel-UHPC composite orthotropic deck bridge, both the field testing and multi-scale finite element analysis modelling were conducted to investigate the fatigue behavior of wrap-around weld at rib-to-floorbeam (RF) joint, and factors influencing stress were further studied to achieve an infinite fatigue life. It is found that a primary tensile stress cycle is jointly produced by the axle group of truck, while a small reversal compressive cycle will be generated when the axle is directly located on the floorbeam. The stress behavior of fatigue detail is extremely sensitive to the localized effect of axle loading instead of entire truck configuration. Since significant stress concentration and high stress gradient exist in the wrap-around weld, it is essential to utilize the hot-spot stress approach rather than nominal stress approach. The wrap-around weld presents an obvious out-of-plane bending deformation, which results from the torsion effect and Poisson’s effect. The Dong’s structural stress reveals that the surface stress is dominated by bending stress. The application of bulkhead can greatly improve the fatigue performance of wrap-around weld. Both the large-scale rib and U-shaped rib are preferable, which are available to achieve an infinite fatigue life without extra fabrication procedure.