The invention discloses an IV-type collagen reagent detection kit. The IV-type collagen reagent detection kit comprises a reagent R1 and a reagent R2. The reagent R1 is prepared from the following components: a Tris buffer solution, sodium chloride, PEG-2000, Arabic gum, trehalose, tween-80, sodium azide, cane sugar, magnesium chloride and water serving as a solvent. The components and concentration of the reagent R2 are as follows: a Tris buffer solution, BSA, glycerinum, an IV type collagen polyclonal antibody, tween-80 and water serving as a solvent. Meanwhile, the invention discloses a preparation method of the reagent detection kit, including preparation of the reagent R1 and the reagent R2. According to the technical scheme disclosed by the invention, the technical defects of high detection cost, complicated detection mode and certain harm to a human body of a radioimmunotherapy adopted in the prior art are successfully overcome. The reagent detection kit disclosed by the invention can be used for rapidly, efficiently and accurately detecting the content of IV-type collagen, so that the rapidness and accuracy of clinical diagnosis are effectively improved.