Abstract:An improved enzyme?linked immunosorbent assay（ELISA）for the detection of tetracycline（TC）was established.BALB/C mice were immunized with the artificial complete antigen conjugating TC and pro?tein，and anti?TC monoclonal antibody（MAb）was then prepared.Acidic potassium permanganate was used to carboxylate the ELISA plate，and TC was coated directly on the plate.The ELISA reaction system was opti?mized to establish an ELISA method employing direct coating of hapten for the detection of TC.The prepared MAb had good specificity and low cross?reaction rate.The established method showed the lowest detection limit IC10 of 0.306 ng/mL，the half inhibition rate IC50 of 9.26 ng/mL，and the recoveries of tetracycline being 96.46%-101.89%and 96.84%-102.50%，respectively，in milk and water.Compared with the method with the coating of artificial antigen（IC10：0.624 ng/mL，IC50：28.24 ng/mL，tetracycline recoveries of 93.86%-105.12% and 94.04%-105.56% in milk and water，respectively），the established method showed signifi?cantly improved detection sensitivity and can be used for the determination of TC content in actual samples，demonstrating a promising application prospect.