Species limits within the clinically important Fusarium incarnatum-F. equiseti and F. chlamydosporum species complexes (FIESC and FCSC, respectively) were investigated using multilocus DNA sequence data. Maximum-parsimony and maximum-likelihood analyses of aligned DNA sequences from four loci resolved 28 species within the FIESC, within which the species were evenly divided among two clades designated Incarnatum and Equiseti, and four species within the FCSC. Sequence data from a fifth locus, beta-tubulin, was excluded from the study due to the presence of highly divergent paralogs or xenologs. The multilocus haplotype nomenclature adopted in a previous study (K. O'Donnell, D. A. Sutton, A. Fothergill, D. McCarthy, M. G. Rinaldi, M. E. Brandt, N. Zhang, and D. M. Geiser, J. Clin. Microbiol. 46:2477-2490, 2008) was expanded to all of the species within the FIESC and FCSC to provide the first DNA sequence-based typing schemes for these fusaria, thereby facilitating future epidemiological investigations. Multilocus DNA typing identified sixty-two sequence types (STs) among 88 FIESC isolates and 20 STs among 26 FCSC isolates. This result corresponds to indices of discrimination of 0.985 and 0.966, respectively, for the FIESC and FCSC four-locus typing scheme using Simpson's index of discrimination. Lastly, four human and two veterinary isolates, received as members of the FIESC or FCSC, were resolved as five phylogenetically distinct species nested outside these species complexes. To our knowledge, these five species heretofore have not been reported to cause mycotic infections (i.e., F. armeniacum, F. brachygibbosum, F. flocciferum, and two unnamed Fusarium species within the F. tricinctum species complex).