To improve the efficiency and accuracy of numerical simulation of single-phase flow and heat transfer in Pressurized Water Reactor (PWR) rod bundle, simulations were performed in two computational models of four-subchannel without spacer and 5×5 rod bundle with a mixing vane spacer under different working conditions. The different turbulence models, near-wall treatments and near-wall mesh generation strategies were comprehensively evaluated from two aspects of heat transfer (Nusselt number) and fluid flow (Secondary flow, turbulence intensity) by using high quality wall-function mesh and low-Reynolds-number mesh. Analysis shows that the numerical results of Nusselt number were seriously affected by both of the turbulence model and near-wall treatment, while secondary flow and turbulence intensity were mainly influenced by turbulence model. The Reynolds Stress Model (RSM) could predict the secondary flow field in bare rod bundle better than eddy viscosity models, and there was no significant difference for calculating the secondary flow field downstream of the mixing vane spacer grid. Among all turbulence models, only the SST k-ω model behaved well even combined with different type of near-wall meshes. Using the SST model with wall-function mesh for engineering computation can provide both the efficiency and accuracy of simulation, and the SST model combined with low-Reynolds-number mesh is recommended for numerical research.