Leaf rust caused by Puccinia triticina f. sp. tritici is the most common and wide-spread rust disease attacking many wheat cultivars in Egypt. Three main methods were used to identify leaf rust resistance genes; gene postulation, genetic analysis and molecular markers. The two resistant wheat cultivars i.e., Giza-171 and Sids-14, as well as the ten monogenic lines for leaf rust resistance; Lr9, Lr19, Lr21, Lr24, Lr25, Lr28, Lr29, Lr34, Lr46 and Lr47, were selected to carry out the present study. Out of the three methods used, genetic analysis and molecular markers were the best to identify the resistance genes in the two studied cultivars. Ten specific primers were used for the identification of 10 resistance genes in the two new Egyptian wheat cultivars i.e., Giza-171 and Sids-14. Six leaf rust resistance genes, Lr9, Lr25, Lr28, Lr29, Lr46 and Lr47 were identified in Giza-171, but only three genes, Lr29, Lr46 and Lr47, were detected in Sids-14. Each of these two new wheat cultivars proved to have an adequate and high level of genetic resistance to leaf rust. The tested wheat cultivars should be used as a good source of leaf rust resistance in breeding programs for rust resistance. Knowledge of the leaf rust resistance genes which has not been designated yet, will help to narrow the gap and throw light on the future objectives for the researchers interested in the full utilization of these genes in breeding materials.