The invention discloses an enzyme-linked immunoassay kit of a structural protein antibody for a seneca valley virus. The kit comprises an elisa plate, positive control serum, negative control serum, an HRP-conjugated antibody, a sample diluent, a 20-fold concentrated detergent, a substrate solution A, a substrate solution B and a stop solution, wherein the elisa plate is coated with a structural protein epitope polypeptide composition for the seneca valley virus. The epitope polypeptide composition is one or any combination of more than two of a polypeptide as shown in a sequence 1, a polypeptide as shown in a sequence 2, a polypeptide as shown in a sequence 3 or a polypeptide as shown in a sequence 4 in the sequence table. The elisa plate is coated with a chemical synthetic antigen peptide, so that the kit is low in antigen dosage and high in sensitivity and specificity, and whether the structural protein antibody is infected by the seneca valley virus or not can be efficiently detected. The kit is high in sensitivity, good in specificity, convenient in operation, and has a good market prospect.