Motion estimation (ME) is one of the most time-consuming parts in video coding. The use of multiple partition sizes in H.264/AVC makes it even more complicated when compared to ME in conventional video coding standards. It is important to develop fast and effective sub-pixel ME algorithms since: 1) the computation overhead by sub-pixel ME has become relatively significant while the complexity of integer-pixel search has been greatly reduced by fast algorithms, and 2) reducing sub-pixel search points can greatly save the computation for sub-pixel interpolation. In this letter, a novel fast sub-pixel ME algorithm is proposed which performs a “rough” sub-pixel search before the partition selection, and performs a “precise” sub-pixel search for the best partition. By reducing the searching load for the large number of non-best partitions, the computation complexity for sub-pixel search can be greatly decreased. Experimental results show that our method can reduce the sub-pixel search points by more than 50% compared to existing fast sub-pixel ME methods with negligible quality degradation.