Clostridium perfringensis the etiological agent for necrotic enteritis (NE) in broiler chickens, which causes a substantial economic loss of an estimated USD 6 billion annually in the global poultry industry. Collagen adhesion is involved in the NE pathogenesis in poultry. In this study, the binding capabilities of chickenC. perfringensisolates of various genetic backgrounds (netB−tpeL−,netB+tpeL−,netB+tpeL+) to collagen types I–V and gelatin were examined, and the putative adhesin proteincnaAgene was investigated at the genomic level. In total, 28C. perfringensstrains from healthy and NE-inflicted sick chickens were examined. The results on collagen adhesin-encoding genecnaAby the quantitative-PCR results indicated thatnetB−tpeL−isolates had much lower copies of the detectablecnaAgene thannetB+isolates (10netB+tpeL−isolates, 5netB+tpeL+isolates). Most of the virulentC. perfringensisolates demonstrated collagen-binding abilities to types I–II and IV–V, while some strains showed weak or no binding to collagen type III and gelatin. However, thenetB+tpeL+isolates showed significantly higher binding capabilities to collagen III thannetB−tpeL−andnetB+tpeL−isolates. The data in this study suggest that the collagen-binding capability of clinicalC. perfringensisolates correlates well with their NE pathogenicity levels, especially forC. perfringensisolates carrying genes encoding crucial virulence factors and virulence-associated factors such asnetB, cnaA, andtpeL. These results indicate that the presence of thecnaAgene may be correlated withC. perfringensvirulence (particularly fornetB+isolates).