The invention discloses border disease virus recombinant E0 protein, application of the E0 protein in the aspect of detecting border disease virus antibodies, and a border disease virus antibody detection ELISA kit. The border disease virus antibody detection ELISA kit includes an ELISA plate containing the BDV recombinant E0 protein serving as coating antigens. The E0 recombinant protein is prepared by cloning gene segments containing main E0 antigen areas into a prokaryotic expression vector pET-32a (+) and obtaining a recombinant gene expression vector; specifically, the E0 recombinant protein is obtained through the following steps that according to a BDV-E0 gene sequence, a pair of specific primers are designed, then, through a RT-PCR method, a E0 gene of the BDV is amplified, the E0gene is directionally inserted into the pET-32a (+) expression vector, screening is conducted to obtain positive recombinant expression plasmids pET-32a (+)-BDV-E0, the plasmids are transformed into BL21 competent cells, and then through ITPG inducible expression, the BDV-E0 recombinant protein is obtained.