【摘要】 通过合成黄曲霉毒素M₁（AFM₁）完全抗原AFM₁-BSA并免疫小鼠,成功制备了AFM₁单克隆抗体2F12.通过以辣根过氧化物酶（HRP）标记的AFB1作为竞争抗原,成功研制了能同时检测液态奶、酸奶和奶粉中AFM₁和AFB1的ELISA试剂盒,该试剂盒的最低检测限（IC10）为37 pg·m L^-1,IC50为211 pg·m L^-1,线性范围为50—1500 pg·m L^-1,对AFM1和AFB1的交叉反应率分别为100%和102%,对其它黄曲霉毒素的交叉反应率小于20%.利用建立的试剂盒对多种样品进行了加标回收实验,回收率在87.9%—109.1%之间,批内相对标准偏差在3.9%—10.8%之间,批间相对标准偏差≤11.1%.多次对比试验表明,试剂盒的检测结果与进口试剂盒相当.更多还原

【Abstract】 A monoclonal antibody 2F12 against AFM₁ was developed by immunizing mice with AFM₁-BSA antigen. With the HRP-labeled AFB₁ as the competitive antigen,an ELISA kit for simultaneous detection of AFM₁ and AFB₁ in milk and milk products was successfully developed. The limit of detection and the IC50 for AFM₁ was 37 pg·m L^-1 and 211 pg·m L^-1,respectively. The linear range was between 50 pg·m L^-1and 1500 pg·m L^-1 for AFM₁,Cross reactivities for AFM1 and AFB1were 100% and 102%,and less than 20% for the other aflatoxin analoges. Spike recoveries for milk and milk products ranged from 87. 9% to 109. 1%,and the intra- and inter-assay relative standard deviations were lower than 10. 8% and 11. 1%. The results of this kit were consistent with that of imported commercial ELISA kits in several tests.更多还原