The invention discloses a reagent kit and method for detecting a thyroglobulin antibody. The reagent kit contains a first reagent, a second reagent, a magnetic separation reagent, a chemical light-emitting substrate, a calibration article, a quality control article and cleaning liquid. The first reagent is a biotin reagent marked by thyroglobulin. The second reagent is a thyroglobulin enzyme combination reagent. The magnetic separation reagent is a superparamagnetic nanometer magnetic particle reagent marked by streptavidine, and the reagent kit is adopted for detecting the thyroglobulin antibody. Due to the mode, the defects that radioimmunoassay is high in pollution, the period of validity is short, common microwell plate enzyme immunoassay precision is poor, and sensitivity is poor are overcome, an alkaline phosphatase enzyme catalysis chemiluminescence system, a magnetic particle separation system and an unique antigen and antibody coupling technology are used, and the sensitivity, the precision, the stability, the validity period, the safety and the environment friendliness of the method are greatly improved.