摘要
为构建快速检测乳制品中黄曲霉毒素M1(AFM1)的直接竞争-酶联免疫吸附(dc-ELISA)体系,将辣根过氧化物酶(HRP)与高纯度抗AFM1单克隆抗体进行偶联后对其与AFM1竞争性反应条件进行优化,并利用AFM1污染标准物质ERMI-BD282(0)、ERMI-BD 283(0.11μg/kg)、ERMI-BD 284(0.44μg/kg)等对检测体系的灵敏度和精确性进行验证。结果当AFM1-BSA包被质量浓度0.25μg/L、抗AFM1 mAb-HRP稀释2000倍时dc-ELISA检测AFM1的IC50为0.75μg/L,检测范围为0.015～4.05μg/L,添加AFM1至0.45μg/L的鲜奶样品中检测回收率平均为80%,dc-ELISA可满足鲜乳中AFM1国家残留限量标准0.5μg/kg的检测要求。该体系可应用于鲜奶制品中AFM 1大于0.5μg/kg污染样品的快速初筛。
A rapid and sensitive direct competitive enzyme-linked immunosorbent assay（ELISA） using anti-AFM1 mAb with horseradish peroxidase（HRP） to measure aflatoxin M1（AFM1） in milk was described.Using AFM1 contaminated milk ERMI-BD282（zero）,ERMI-BD 283（0.11 μg/kg） and ERMI-BD 284（0.44μg/kg）,the sensitivity and accuracy of the developed assay was validated.The optimized assay conditions regarding sensitivity and stability were found to be： coating AFM1-BSA antigen concentration 0.25 μg/kg and dilution factor of anti-AFM1 mAb-HRP conjugate 2000.Assays of ERMI-BD282 samples spiked with AFM1 at the level of 0.45 μg/L revealed an average recovery of around 80%.The developed method showed an IC50 of 0.75μg/L and a linear range of 0.015-4.05 μg/L.This assay may be used in rapid screening of contaminated milk at AFM1 0.5μg/L.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2011年第18期221-224,共4页
Food Science
基金
黑龙江省留学归国人员项目(Lc05c11)
