The invention discloses a method for detecting an anti-insulin antibody in human serum. The method comprises the following steps: preparing a cell carrier; preparing other components; screening positive serum expressing the anti-insulin antibody and preparing a positive control liquid; preparing a kit, wherein the kit comprises the cell carrier used for expressing the anti-insulin antibody, a negative control liquid, the positive control liquid, an enzyme substrate, a dilute liquid and a stop liquid; finally, performing carrier expression on the anti-insulin antibody in the diluted positive control liquid and the human serum for experiment, and carrying out analyzing. According to the method, the positive control liquid is diluted and subjected to data selection, and the cell carrier is expressed by selecting the numerical range of intensively dispersed data, so that the problem of an instable result caused by an instable positive control antibody in each batch during detection of the anti-insulin antibody content of the human serum by the conventional kit is solved.