The present invention provides a cocktail antigen for bovine tuberculosis allergic reaction detection, comprising ESAT6 antigen, CFP10 antigen, FixB antigen and HSPX antigen; wherein, the mass ratio of ESAT6 antigen, CFP10 antigen, FixB antigen and HSP antigen is 2:2 :1.5:1.5. The cocktail antigen provided by the invention can be used to prepare the detection antigen for bovine tuberculosis allergic reaction. The cocktail antigen of the present invention can replace bovine-type PPD and poultry-type PPD used in comparative allergy, and the effect of comparative allergy can be achieved by using the cocktail antigen for a single intradermal allergy. , After 72 hours, return to the field again to measure the skin thickness, calculate the skin thickness difference and other series of operations, simplified into a single point operation without calculation, the whole detection process is more convenient. At the same time, non-specific mycobacterial infections such as mycobacterium avium can be excluded, and bovine tuberculosis can be diagnosed more specifically.