Stripe rust is caused by Puccinia striiformis f. sp. tritici (Pst) is threatening wheat production in Ethiopia. Wheat cultivars succumb to new Pst race (s) soon after their release from research centers. The objective of this study was to determine stripe rust resistance in selected Ethiopian commercial and elite bread wheat lines. In 2017, a total of 37 commercial and elite bread wheat lines were exposed to the prevailing stripe rust races in hot spot areas (Kulumsa and Meraro) in Arsi zone of Oromia region. In the second year (2018), only promising lines 22 (16 commercial bread wheat and 6 elite’s lines) were evaluated both at seedling and adult plant growth stages. The seedling test was conducted in the greenhouse at Kulumsa research center using three Pst races. Of the 37 commercial and elite bread wheat lines, 19 (51.4%) exhibited lower or equal disease reaction compared to the resistant check (Enkoy) across locations and seasons. Eleven bread wheat lines showed both adult plant and seedling resistance. The 37 commercial wheat lines that showed field resistance further exposed to three Pst races at seedling stage and 11 exhibited seedling resistance to all races. This study has identified seedling and adult plant resistance in the commercial and elite bread wheat lines to the prevailing Pst races.