Indirect ELISA Using Multi–Antigenic Dominants of p30, p54 and p72 Recombinant Proteins to Detect Antibodies against African Swine Fever Virus in Pigs

African swine fever (ASF) caused by ASF virus (ASFV) is a fatal disease in pigs and results in great economic losses. Due to the lack of available vaccines and treatments, serological diagnosis of ASF plays a key role in the surveillance program, but due to the lack of knowledge and the complexity of the ASFV genome, the candidate target viral proteins are still being researched. False negativity is still a big obstacle during the diagnostic process. In this study, the high antigenic viral proteins p30, p54 and p72 were screened to find the antigenic dominant domains and the tandem His–p30–54–72 was derived. An indirect enzyme–linked immunosorbent assay (iELISA) coated with His–p30–54–72 was developed with a cut–off value of 0.371. A total of 192 clinical samples were detected by His–p30–54–72–coated indirect ELISA (iELISA) and commercial ASFV antibody kits. The results showed that the positive rate of His–p30–54–72–coated iELISA was increased by 4.7% and 14.6% compared with a single viral protein–based commercial ASFV antibody kits. These results provide a platform for future ASFV clinical diagnosis and vaccine immune effect evaluation.