The invention discloses a two-in-one determination kit for serum amyloid protein A and C-reactive protein. A test strip is arranged on the determination kit, and the determination kit is characterized in that a PVC board, a sample pad, a combined pad, a nitrocellulose membrane and a water absorbent pad are sequentially arranged on the test strip from bottom to top, wherein two monoclonal antibody-micro-spherical coupling compounds of the anti-serum amyloid protein A and anti-C-reactive proteins which are respectively marked by rare earth Eu<3+> fluorescent microspheres are adsorbed on the combined pad; the diameter of the rare earth fluorescent microspheres is 150nm; the fluorescent microspheres contain rare earth lanthanide elements Eu<3+> and is stable in a basic state, and fluorescence with wavelength of 615nm can be emitted under an action of an excitation light source with the length of 337nm; the monoclonal antibodies are the purified and mixed monoclonal antibodies, which are respectively originated from monoclonal antibody cell strains for 2-6 different serum amyloid protein A and C-reactive protein antigenic epitopes. The two-in-one determination kit has the advantages of convenient operation, quick reaction, high sensitivity, strong specificity and the like.