Abstract: The quantitative detection of Aflatoxin B1(AFB1) was investigated on the microsphere array platform and indirect competition by using the Anti-Aflatoxin B1.Through optimizing the concentration and saturated concentration of AFB1-BSA and Anti-Aflatoxin B1 as well as incubation time of antigen and antibody,the method of quantitative detection of AFB1 was established.As referring to the detection sensitivity standard IC50,results showed that the detection sensitivity was 1.33 ng/mL.Meanwhile,as referring to the limit of detection standard IC10,the detection limition was 0.15 ng/mL.The linear equation was y=0.000 6x+0.000 8.By applying this original research to test the imitating contaminated skim and full milk,the recovery rate of the two kinds of milk was all greater than 75% under the AFB1 added level of 2.0-16.0 ng/mL,and the relative standard deviation was between 2.80%-4.69%(n=7).Overall,the quantitative analysis of AFB1 using the microsphere array technology was sensitive,stable,rapid and simple.The method was applicable to mass sample testing.