The invention relates to the technical field of biotechnology, and specifically relates to a p16 immune cell chemical labeling color-developing kit. The detection kit comprises the following reagents:0.5-1.0 ml of a p16 immune cell standard substance with the concentration of 81 pg/mL, 1.5 to 2.0 ml of a standard substance diluent, 2-4 ml of horseradish peroxidase, 2-4 ml of a sample diluent, 2-4ml of a color-developing agent solution A, 2-4 ml of a color-developing agent solution B, 2-4 ml of a stop solution, and 20ml of 20 times of concentrated washing solution. The method comprises the steps of coating a microporous plate with a purified human p16 antibody to prepare a solid-phase antibody, adding p16 into micropores coated with a monoclonal antibody, combining with a horseradish peroxidase-labeled p16 antibody to form an antibody-antigen-enzyme-labeled antibody compound, thoroughly washing, and then adding a color-developing agent A substrate for color development. The color-developing agents A and B are converted into blue under the catalysis of horseradish peroxidase and converted into final yellow under the action of acid, the positioning effect and the color-developing effect are good, the labeling is more obvious, the color depth is in positive correlation with p16 in a sample, and the absorbance is measured by using a microplate reader at the wavelength of 450 nm.