The invention discloses an African swine fever virus synthetic peptide ELISA antibody detection kit. The kit includes an enzyme-labeled plate, a positive control serum, a negative control serum, an enzyme-labeled secondary antibody, a sample dilution solution, a 20-fold concentrated washing solution, a substrate solution A, a substrate solution B and a stop solution. The enzyme-labeled plate is coated with African swine fever virus epitope polypeptide composition. The epitope polypeptide composition is any combination of one or two or more of a polypeptide shown as a sequence 1 in a sequence listing, a polypeptide shown in a sequence 2 in the sequence listing and a polypeptide shown as a sequence 3 in the sequence listing. The kit uses a chemically synthesized antigen peptide-coated enzyme-labeled plate with low antigen consumption, high sensitivity and specificity, and can efficiently detect whether African swine fever virus antibodies exist or not. The kit is high in sensitivity, good in specificity, convenient to operate and good in market prospects.