The invention discloses a thyrotropic hormone quantitative detection kit which comprises a magnetic particle suspension coated with an antithyrotropichormone monoclonal antibody, a thyrotropic hormone serial calibration product, a antithyrotropichormone monoclonal antibody marked by horse radish peroxidase, a luminous substrate solution A, a luminous substrate solution B and a concentrated wash solution. The invention also discloses a preparation method of the kit. The invention has the advantages that a double-antibody-sandwich one-step reaction mode is adopted to preferentially select a pair of monoclonal antibodies with higher specificity and stronger appetency from a large quantity of active materials, and the monoclonal antibodies are respectively used as a coated antibody and an enzyme marked antibody, so that the diagnosis sensitivity, the precision and the detection range of the kit are obviously superior to that of like kits, and the reaction time is also greatly shortened; meanwhile, an antithyrotropic hormone monoclonal antibody and immune magnetic particle coupling method is adopted, the coupling efficiency is higher, the bonding is firm and the process is stable, thusthe product cost is greatly reduced while the product properties are improved.