摘要
目的建立高灵敏度的时间分辨免疫法检测黄曲霉毒素B_1的含量。方法通过双功能螯合剂异氰酸苄基二乙烯三胺四乙酸络合稀土元素Eu^(3+)标记抗AFB_1的单克隆抗体,以AFB_1-OVA为固相抗原,优化反应条件,建立直接竞争时间分辨免疫检测方法。结果优化条件后,方法灵敏度为0.02μg/L,IC_(50)为0.73μg/L,线性检测范围为0.01~30μg/L,大米、花生、黄豆和干果样品回收率在91%~104%之间,检测结果准确;与结构类似物AFB_2、AFG_1、AFG_2、AFM_1的交叉反应率分别为17.3%、4.49%、2.37%和0.73%。结论本方法灵敏度高、特异性强、检测快速,可以满足实际样品的检测需求。
Objective To establish a hypersensitive time-resolved fluoroimmunoassay （TRFIA） method for detection of aflatoxin B1 （AFB1）. Methods The antibody for anti-AFB1 was labeled by Eu3^＋-DTTA, AFB1-OVA was bounded on the surface of a microtiter plate, the reaction condition was optimized and then the direct competition with TRFIA was established. Results Under optimized assay conditions, the limit of detection was 0.02 μg/L, the IC50 of TRFIA was 0.73μg/L, the linear detection range was 0.01-30 μg/L, the recoveries for rice, peanuts, soybeans and preserved fruits were range from 91% to 104%, which showed the results of detection were accuracy; and the cross reaction with AFB2, AFG1, AFG2 and AFM1 were 17.3%, 4.49%, 2.37% and 0.73%, respectively. Conclusion The established method is sensitive, specific and rapid, which can meet the detection demand of actual samples.
出处
《食品安全质量检测学报》
CAS
2017年第3期796-802,共7页
Journal of Food Safety and Quality
