The invention provides a colloidal gold test strip detection card capable of simultaneously detecting a bovine brucellosis antibody and bovine tuberculosis gamma-interferon. The colloidal gold test strip detection card comprises a carrier plate, a sample pad, a gold label pad, a nitrocellulose membrane and a water absorption pad, wherein the sample pad and the water absorption pad are located at the two ends of the carrier plate; the nitrocellulose membrane is located in the middle of the carrier plate; the gold label pad loaded with a gold-labeled anti-gamma-interferon monoclonal antibody anda gold-labeled rabbit anti-bovine secondary antibody is arranged on the junction of the sample pad and the nitrocellulose membrane; the edge of one end of the gold label pad is overlapped under the sample pad, and the edge of the other end of the gold label pad is overlapped on the nitrocellulose membrane; a bovine tuberculosis detection line, a brucellosis antibody detection line and a quality control line are arranged from the side, close to the gold label pad, of the nitrocellulose membrane; the bovine tuberculosis detection line is coated with an anti-gamma-interferon monoclonal antibodyMT17.1, the brucellosis antibody detection line is coated with brucella lipopolysaccharide antigens, and the quality control line C is coated with goat anti-mouse IgG. The detection card can be used for effectively detecting bovine brucellosis and bovine tuberculosis.