Here, we describe two novel multigene families (MGF) present in the terminal variable regions of the African swine fever virus (ASFV) genome. MGF530 includes at least six related ORFs (averaging 530 amino acids) containing four well-conserved domains and amino acid identities ranging from 24 to 55%. MGF300 is composed of three ORFs (averaging 300 amino acids) containing three highly conserved domains and amino acid identities ranging from 25 to 46%. Amino terminal regions of predicted MGF530 and 300 proteins share significant similarity with each other and with the comparable regions of previously characterized MGF360 ORFs. Members of both MGF530 and 300 are transcribed in ASFV-infected porcine macrophages. These results indicate that the composition and organization of MGF in the terminal variable region of the ASFV genome are more complex than previously recognized.