The invention discloses an immunofluorescence detection test strip and a preparation method thereof for rapid quantitative detection of porcine epidemic diarrhea viruses. The test strip comprises a sample cushion, a combination cushion, a chromatography film and a water-absorbing cushion, wherein the combination cushion is provided with a fluorescent microsphere labelled anti-PEDV (Porcine Epidemic Diarrhea Viruses) single-domain antibody; the single-domain antibody has high specificity and high sensibility against the antigen PEDV. The test strip is used for detecting PEVD viruses in breeding pig manure or PEDV pollution in feeding stuff plasma proteins on the basis of the newfound immunology principle of the specific single-domain antibody and antigen, can be used for on-site rapid detection or laboratory detection by simply taking 5-10 min; when the test strip is used in combination with a fluorescence quantitative detector, quantitative detection can be realized; the test strip is simple and convenient to operate, operators don't need professional training, a special laboratory is not needed, the limitation of the conventional detection method is overcome, and the test strip has good market prospects.