Development of African Swine Fever Virus Pseudotype Systems for Ba71v and Georgia 2007 strains

African swine fever virus (ASFV) is a large, complex DNA virus that causes a severe haemorrhagic disease in domestic swine and Eurasian wild boar. Due to its high pathogenicity, research involving ASFV is restricted to high-containment laboratories, which significantly limits investigations into its biology, host interactions and the development of effective control measures. To overcome these limitations, this study utilised pseudotyped virus (PV) technology to identify ASFV proteins involved in cell entry. Candidate proteins were identified through bioinformatic analysis and a review of the published literature, incorporated into PVs, and subsequently assessed for their ability to facilitate cellular entry. This approach identified a combination of three ASFV proteins that, when combined, resulted in the most efficient cell entry. This work represents the successful generation of the first functional ASF pseudotype system for both Ba71v and Georgia 2007 strains, enabling the safe study of ASFV entry mechanisms under containment level 2.