The invention relates to a kit for detecting urine transferrin through a chemiluminescence enzyme immunoassay method, and a preparation method thereof. The kit comprises an antibody-coated plate, a urine transferrin enzyme antibody, a coating buffer solution, a blocking solution, a urine transferrin standard substance, a standard substance dilution solution, a washing solution, a sample dilution solution and a chemiluminescent substrate solution. According to the present invention, the kit can detect the UTF concentration level of human based on chemiluminescence enzyme immunoassay; the chemiluminescent substrate catalytic enzyme HRP and the anti-UTF antibody are conjugated, the obtained conjugate reacts with the sample or the antigen to form the immune complexes, the chemiluminescent substrate solution is added, the relative light unit of the chemiluminescent reaction is measured, the UTF antigen content in the standard substance/the sample is proportional to the relative light unit measured by the optical system, and through standard curve fitting, the UTF content in the urine sample is determined; and the kit of the invention has advantages of high sensitivity, strong specificity, easy operation, low cost and the like.