Abstract: The present disclosure provides a test kit for simultaneously detecting a plurality of analytes, in which the test kit includes a lysis solution and a test strip. The lysis solution includes a salt, a surfactant, a stabilizer, and a buffer solution. The test strip includes a sample pad, a conjugation pad, a cellulose membrane, and a water-absorbing pad sequentially arranged on a support plate. The conjugation pad includes a conjugation pad solution and a plurality of antibody-conjugated microspheres, and the antibody-conjugated microspheres recognize plurality of analytes. The test kit of the present disclosure achieves an effect of simultaneously detecting more than or equal to four analytes even though the conjugation pad has a limited capacity of the antibody by preparing the lysis solution with appropriate ingredients and improving a formulation of the solution contained in the test strip.