The invention discloses a chemiluminescence quantitative detection kit for estradiol. The kit comprises an estradiol detection reaction plate, an enzyme conjugate, a luminescent substrate, a calibrator, a quality control material and washing concentrate, wherein the reaction plate is coated with an estradiol antibody. The kit can specifically and quantitatively detect the content of the estradiol in patient serum, and is used for the auxiliary diagnosis of diseases such as ovarian hypoplasia, primary ovarian failure, pituitary amenorrhea or infertility, Cushing's syndrome, Addison's disease, malignant tumors, focal lesion of brain and hypophysis, and the like. Compared with the conventional enzyme linked immunosorbent assay (ELISA) technology, the chemiluminescence immunoassay keeps high specificity of the ELISA technology, stability and reliability of a detection result, and convenience of operation, and can improve detection sensitivity simultaneously.