Spot blotch, caused by Cochliobolus sativus, is an economically important disease of barley. To identify genetic loci conferring resistance to three different pathotypes of C. sativus, a worldwide barley core collection (BCC) consisting of 1480 accessions from the USDA National Small Grains Collection were genotyped with the barley 9k Illumina Infinium iSELECT assay and phenotyped at the seedling stage with three C. sativus isolates ND85F (pathotype 1), ND90Pr (pathotype 2), and ND4008 (pathotype 7). Association mapping analysis was performed with the Whole_Panel containing 1480 barley accessions, as well as Two-rowed_Panel and Six-rowed_Panel consisting of 621 two-rowed and 857 six-rowed barley accessions, respectively. For resistance to isolate ND4008, one quantitative trait locus (QTL, QRcs-6H-P7) was detected in all three panels. Three other QTL (QRcs-1H-P7, QRcs-2H-P7, and QRcs-3H-P7) were detected in Whole_Panel, Six-rowed_Panel, and Two-rowed_Panel, respectively. For resistance to isolate ND90Pr, one QTL (QRcs-1H-P2) was identified in the Whole_Panel and the Two-rowed_Panel, and the other QTL (QRcs-6H-P2) was only identified in the Six-rowed_Panel. For resistance to isolate ND85F, three QTL (QRcs-1H-P1, QRcs-3H-P1, QRcs-7H-2-P1) were detected in all three panels, and one QTL (QRcs-7H-1-P1) was only detected in the Two-rowed_Panel. Among the ten QTL detected, four (QRcs-1H-P1, QRcs-3H-P1, QRcs-7H-2-P1, and QRcs-1H-P2) were mapped to chromosome regions containing previously identified QTL for spot blotch resistance, while six (QRcs-1H-P7, QRcs-2H-P7, QRcs-3H-P7, QRcs-6H-P7, QRcs-6H-P2, and QRcs-7H-1-P1) were novel. The SNP markers associated with the QTL identified in this study will be useful for breeding barley cultivars with resistance to multiple pathotypes of C. sativus.