The invention relates to the technical field of biology and in particular relates to a kit for simultaneously detecting an avian leukosis virus antibody and a salmonella pullorum antibody. An antigencombination of the kit comprises p27 protein, gp85 protein and GroEL-delta8-1 protein. Avian leukosis virus capsid protein p27, prokaryotic expression protein of envelope protein gp85, and prokaryoticexpression protein of truncated GroEL-delta8-1 of a salmonella pullorum dominant antigen are used as a coating antigen to develop an ELISA (Enzyme-linked Immunosorbent Assay) kit capable of simultaneously detecting the avian leukosis virus antibody and the salmonella pullorum antibody. Compared with a current common kit for independently detecting avian leukosis or pullorum disease, the kit provided by the invention has the effect of simultaneously detecting the avian leukosis or the pullorum disease, and the detection and purification work of the avian leukosis and the pullorum disease can be extremely alleviated.