Provided in the present invention is a whole blood one-step alpha-fetoprotein colloidal gold detection kit, comprising a kit seat, a kit lid and a reagent strip, wherein the reagent strip is fixed between the kit seat and the kit lid, the kit lid is equipped with an observation hole, and the side between the kit lid and the kit seat is equipped with a sampling hole, and the kit can be used for drawing blood directly from the fingertip or for sampling detection. The reagent strip is divided into an upper layer and a bottom layer, the bottom layer is a plastic film, and the upper layer is a glass fiber film, a nitrocellulose film I, a plastic protective film and a nitrocellulose film II successively arranged head-to-tail. The glass fiber film is equipped with rabbit IgG colloidal gold, the nitrocellulose film I is a bridge film equipped with alpha-fetoprotein colloidal gold, and the nitrocellulose film II is equipped with an anti-alpha-fetoprotein colloidal monoclonal antibody and a goat anti-rabbit IgG.
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