In this paper, Eulerian two-phase model coupled with wall boiling model was employed to simulate the departure from nucleate boiling (DNB) in a 5 × 5 fuel assembly. The numerical simulation accurately predicted the critical heat flux (CHF) and also its location. Compared with the calculated results from W-3 formula and FLICA subchannel code, the CFD (Computational Fluid Dynamics) calculations showed good agreement against experimental measurements with the deviation of 5%, but presented more detailed flow distribution. Based on CFD results, the method of CHF prediction was discussed with an eye on wall superheat, void fraction and vapor convective heat flux in rod surfaces. The distribution of flow field and liquid–vapor fraction affected by mixing vane were further studied. Mixing vane can continually influence the downstream thermal hydraulic characteristics, forming the diagonal circulation and square circulation with the flow development. As a result, the bubble aggregation firstly occurred on the surface of corner hot rod instead of central hot rod, which induced the local heat transfer crisis. The CFD calculations presenting the evolution process of physical parameters can provide guidance for rod bundle CHF test and fuel assembly design.