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高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用

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孙清, 李谷丰, 邓乾民, 刘杰民, 时国庆. 高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用[J]. 环境化学, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
引用本文: 孙清, 李谷丰, 邓乾民, 刘杰民, 时国庆. 高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用[J]. 环境化学, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
shu
SUN Qing, LI Gufeng, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of a highly sensitive ELISA kit for aflatoxin B1[J]. Environmental Chemistry, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
Citation: SUN Qing, LI Gufeng, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of a highly sensitive ELISA kit for aflatoxin B1[J]. Environmental Chemistry, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
shu

高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用

  • 1.

    北京科技大学土木与环境工程学院, 北京, 100083;

  • 2.

    北京科技大学化学与生物工程学院, 北京, 100083;

  • 3.

    北京普赞生物技术有限公司, 北京, 100085

  • 基金项目:

    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(FRF-TP-12-007B)资助.

Development and application of a highly sensitive ELISA kit for aflatoxin B1

  • 1.

    School of Civil and Environmental Engineering, University of Science and Technology Beijing, Beijing, 100083, China;

  • 2.

    School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, 100083, China;

  • 3.

    Beijing Primebiotek Company Limited, Beijing, 100085, China

  • Fund Project:

  • 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0.2 μg·mL-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·mL-1, 检测限为7.6 pg·mL-1, 检测线性范围为10—810 pg·mL-1.试剂盒对不同AFB1添加水平(0.5 μg·kg-1, 1 μg·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.
    Abstract: 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 was 7.6 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.
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  • 收稿日期:  2015-03-10
  • 刊出日期:  2015-10-15
孙清, 李谷丰, 邓乾民, 刘杰民, 时国庆. 高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用[J]. 环境化学, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
引用本文: 孙清, 李谷丰, 邓乾民, 刘杰民, 时国庆. 高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用[J]. 环境化学, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
shu
SUN Qing, LI Gufeng, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of a highly sensitive ELISA kit for aflatoxin B1[J]. Environmental Chemistry, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
Citation: SUN Qing, LI Gufeng, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of a highly sensitive ELISA kit for aflatoxin B1[J]. Environmental Chemistry, 2015, 34(10): 1845-1853. doi: 10.7524/j.issn.0254-6108.2015.10.2015031001
shu

高灵敏黄曲霉毒素B1酶联免疫试剂盒的研制及应用

  • 1.  北京科技大学土木与环境工程学院, 北京, 100083;
  • 2.  北京科技大学化学与生物工程学院, 北京, 100083;
  • 3.  北京普赞生物技术有限公司, 北京, 100085
  • 收稿日期:  2015-03-10
基金项目:

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(FRF-TP-12-007B)资助.

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0.2 μg·mL-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·mL-1, 检测限为7.6 pg·mL-1, 检测线性范围为10—810 pg·mL-1.试剂盒对不同AFB1添加水平(0.5 μg·kg-1, 1 μg·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.

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