Fusarium head blight (FHB), primarily caused by Fusarium graminearum Schwabe [telemorph: Gibberella zeae Schw. (Petch)], can significantly reduce the grain quality of wheat (Triticum aestivum L.) due to mycotoxin contamination. Two US soft red winter wheat cultivars, Bess and NC‐Neuse, have moderate resistance to FHB. The objective of this study was to validate genomic regions associated with FHB resistance identified in previous studies involving NC‐Neuse and the cultivar Truman, a full‐sib of Bess. A total of 98 doubled haploid lines derived from the cross Bess × NC‐Neuse were evaluated in inoculated, mist‐irrigated field nurseries. The lines were evaluated for FHB incidence, severity, Fusarium‐damaged kernels, and deoxynivalenol content in seven environments between 2011 and 2014. A 3338‐cM linkage map was developed based on 4014 simple sequence repeat and single nucleotide polymorphism markers. Twelve quantitative trait loci (QTL) associated with FHB resistance were identified. NC‐Neuse alleles provided resistance at QTL on five chromosomes and Bess alleles provided resistance at QTL on five other chromosomes. Alignment of linkage maps revealed that five of these QTL were overlapping with previously identified regions. Quantitative trait loci on chromosomes 1A, 4A, and 6A identified in this study overlapped with QTL regions identified in NC‐Neuse, and QTL identified on chromosomes 2B and 3B overlapped with QTL regions identified in Truman. A preliminary test using Kompetitive Allele‐Specific polymerase chain reaction assays on recent Uniform Southern Winter Wheat Scab Nursery entries showed that the assays developed for Qfhb.nc‐2B.1 may be good candidates for use in marker‐assisted selection.