The set of 44 near-isogenic lines of spring wheat cv. Thatcher and wheat genotypes known to carry specific leaf rust resistance genes were included in the present study for the preliminary validation of twelve STS and SCAR markers linked to leaf rust resistance genes Lr9, Lr19, Lr20, Lr21, Lr24, Lr25, Lr26, Lr28, Lr29, Lr37. Seven Lr genes were specifically tagged by STS and SCAR markers. The presence of genes Lr9, Lr19, Lr20, Lr24, Lr28, Lr29, Lr37 in the tested plant materials was confirmed by a unique amplification of markers SCS5550, SCS265512 and SCS253736, STS638, SCS73719, SCS421570, IPY10 and Lr29F24/R24, cslVrgal3, PS10R/L, respectively. Evaluation of the repeatability and the reliability of selected markers (pTAG621 for Lr1, STS683 for Lr20, D14L for Lr21, Lr25F20/R19 for Lr25, Lr29F24/R24, IPY10 for Lr29, cslVrgal3 for Lr37 and PS10R/L for Lr47) across four European laboratories and PCR conditions demonstrated the usability of STS638, Lr29F24/R24, IPY10, cslVrgal3 and PS10R/L markers in marker-assisted selection. STS markers pTAG621 for gene Lr1, D14L for gene Lr21, Lr25F20/R19 for gene Lr25 were found to be unsuitable for effective screening of large segregating populations in breeding programs.