Abstract: Objective To develop a sensitive,specific and rapid enzyme-linked immunosorbent assay for detection of total aflatoxins.To develop rapid detection EL ISA-kit that possessing country patent.Methods A rapid,quantitative EL ISA-based method(EL ISA-kit)were developed on the base of monoclonal antibodies against Aflatoxins.Results For Aflatoxin(B+G),the limited concentration of detection of the EL ISA-kit was 0.26 ng/ml,linear range was 0.26～20.00 ng/ml,the linear equation was y=-0.4463x+0.3532(R²=0.9915).The inhibition concentration of 50% for Aflatoxin(B+G)was 2.08 ng/ml.The coefficients of variation was 4.18% within the strips,and 5.21% among the level of 0.30μg/kg,2.10μg/kg and 10.40% was 98.63%,94.56% and 112.05% respectively.For peanut recovery rate on the level of 0.50μg/kg,2.00μg/kg and 4.00μg/kg was 88.66%,87.50% and 85.60% respectively,The kit could be stored at 37℃ over 96 hours.The cross reaction rate with aflatoxin B1,B2,G1,G2 was 100% 57.5%,104%,19% respectively,and no cross reaction with other mycotoxins.Conclusion The EL ISA-Kit for total aflatoxins is developed.