摘要
为实现对环境及食品样品中黄曲霉毒素B1的高灵敏检测,通过优化一系列试剂盒参数,研制了一步间接竞争ELISA检测试剂盒.优化后的包被缓冲液为90 mmol·L-1、pH 4.6的柠檬酸缓冲液,最佳反应pH值为7.4,抗体包被浓度为0.2μg·m L-1,HRP-BSA-AFB1稀释比为1/4000,标品稀释液为含7%甲醇的PBST溶液.优化后试剂盒IC50值为66 pg·m L-1,检测限为7.6 pg·m L-1,检测线性范围为10—810 pg·m L-1.试剂盒对不同AFB1添加水平(0.5μg·kg-1,1μg·kg-1)的玉米、豆粕和鱼粉样品平均回收率为108.4%—134.8%.对玉米、豆粕和鱼粉样品各20份盲样测试结果表明,试剂盒检测结果与HPLC-MS/MS检测结果吻合.
A one-step indirect competitive ELISA kit for AFB1 was successfully developed,for the detection of trace AFB1 in environment and foodstuffs.Optimization was perfonmed for coating buffer,pH of assay buffer,concentrations of anti-AFB1 monoclonal antibody and HRP labeled AFB1-BSA,and sample dilution reagent. The limit of detection and the IC50 for AFB1 was7. 6 pg·mL^-1 and 66 pg·mL^-1 respectively,with good linearity between 10—810 pg·mL^-1 for AFB1. Spike recoveries for maize,soybean meal and fishmeal samples ranged from 108. 4% to 134.8%. Aflatoxin B1 residues in sixty samples were determined by the ELISA kit and high performance liquid chromatography-tandem mass spectrometry（ HPLC-MS / MS） respectively,and the result of ELISA kit was consistent with that of HPLC-MS / MS.
出处
《环境化学》
CAS
CSCD
北大核心
2015年第10期1845-1853,共9页
Environmental Chemistry
基金
国家高技术研究发展计划(2012AA101609-5)
新世纪优秀人才支持计划(NCET-11-0581)
中央高校基本科研业务费专项资金(FRFTP-12-007B)资助
