The invention relates to a leptin detection kit. The leptin detection kit comprises a leptin reference product, an antibody-coated microtiter plate, an enzyme-labeled anti-leptin, a washing solution, a color development solution and a stop solution, wherein the leptin reference product is of a recombinant human leptin polypeptide antigen, and the antibody-coated microtiter plate and the enzyme-labeled anti-leptin are of recombinant human leptin polypeptide monoclonal antibodies. The detection principle of the leptin detection kit disclosed by the invention is as follows: the recombinant human leptin polypeptide monoclonal antibody is taken as the antibody-coated microtiter plate, a serum sample to be detected is added, the corresponding enzyme-labeled antibody is further added, a compound of the coating antibody, the serum sample to be detected and the enzyme-labeled antibody is generated, the color development solution is added for color development, then the stop solution is further added for stopping, the residual solution is washed away by the washing solution, the light absorption value of the serum sample to be detected is determined, and the leptin content in the serum sample to be detected can be obtained by comparing with a standard curve.