The invention discloses a lateral flow immunochromatographic determination product for detecting yellow fever viruses and a preparation method of the lateral flow immunochromatographic determination product. The lateral flow immunochromatographic determination product for detecting yellow fever viruses provided by the invention comprises a sample pad, a water sucking pad and a coating film which are connected mutually, wherein the coating film is provided with a detecting line and a quality control line which are separated mutually, and the coating film is located between the sample pad and the water sucking pad. The lateral flow immunochromatographic determination product for detecting yellow fever viruses is characterized in that a magnetic nanoparticle marked yellow fever virus antibody is loaded on the sample pad, the detecting line is coated with a yellow fever virus coating antibody, and the quality control line is coated with a second antibody specifically combined with the magnetic nanoparticle marked yellow fever virus antibody; and the magnetic nanoparticle marked yellow fever virus antibody is a polymer formed by the peptide bond covalent binding of a yellow fever virus antibody to be marked and carboxyl modified magnetic nanoparticles. The experiments of the invention show that the product for detecting yellow fever viruses provided by the invention is high in sensitivity, strong in specificity, rapid and convenient, and can realize objective determination.