Gene stability assessment of ASFV-GZdeltaI177LCD2vMGF in vitro and in vivo. To this end, the vaccine virus was mixed with the same dose of wild strain in equal volume (104.0 TCID50 in 500ul), and then infected BMDM cells at a multiple of infection (m.o.i) of 0.01. After 72 h post-infection, the presence of fluorescence was detected under a fluorescence microscope (ECHO Global) and the proportion of infected cells was calculated. The virus culture was harvested for four more passages in BMDM cells at a 2 % (V/V) dose. we purified the fifth-generation virus by plaque purification and limiting dilution, followed by nanopore sequencing to evaluate its gene stability.