The invention relates to a fluorescent microsphere detection device for gastric function and gastric cancer risk and a preparation method thereof, and belongs to the field of medical detection equipment. The device is prepared by adhering a nitrocellulose membrane with solid phase containing high-specificity gastrin 17, pepsinogen I, pepsinogen II antibody, anti-human IgG antibody, anti-human IgMantibody and goat-anti-mouse IgG polyclonal antibody, glass fibers adsorbed with fluorescent microsphere labeled gastrin 17, pepsinogen I, pepsinogen II antibodies and helicobacter pylori urease antigens, a sample pad, absorbent paper and other auxiliary materials. The reaction sensitivity is effectively improved; under the same threshold value, the dosage of the immunofluorescence microspheres can be reduced, the cost is saved; five gastric cancer risk markers including gastrin 17, pepsinogen I, pepsinogen II, helicobacter pylori urease IgG antibody and helicobacter pylori urease IgM antibodyin a specimen can be detected at the same time, and the complexity of production operation is not increased. The detection test paper is high in sensitivity, strong in specificity and strong in practicability.