The invention discloses an enzyme-linked immunosorbent assay kit for detecting diethoxy phosphorothioate organic phosphorus pesticide, which comprises a kit body, an ELISA plate coated with the antigen of the diethoxy phosphorothioate organic phosphorus pesticide, an antibody solution against diethoxy phosphorothioate organic phosphorus pesticide, a second anti-solution, a standard solution, a concentrated extracting solution, a substrate solution, a buffer solution of the substrate, a concentrated washing liquid and a stopping liquid. The organic phosphorus pesticide refers to the organic phosphorus pesticide containing the common structure of diethoxy phosphorothioate, in particular parathion, coumaphos, quinalphos, triazophos, phoxim, dichlofenthion, phorate, chlorpyrifos, bromophos ethyl, isazofos, diazinon, sulfotep and a mixture thereof. The enzyme-linked immunosorbent assay kit can be used to detect the organic phosphorus residues in fruits, vegetables, soil and water, and isconvenient in operation, high in sensitivity, low in price and is suitable for high-throughput screening monitored on spot.