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黄曲霉毒素M1和黄曲霉毒素B1总量酶联免疫试剂盒的研制及应用

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孙清, 李馥雪, 贾玉军, 邓乾民, 刘杰民, 时国庆. 黄曲霉毒素M1和黄曲霉毒素B1总量酶联免疫试剂盒的研制及应用[J]. 环境化学, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
引用本文: 孙清, 李馥雪, 贾玉军, 邓乾民, 刘杰民, 时国庆. 黄曲霉毒素M1和黄曲霉毒素B1总量酶联免疫试剂盒的研制及应用[J]. 环境化学, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
shu
SUN Qing, LI Fuxue, JIA Yujun, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of an ELISA kit for simultaneous determination of aflatoxin M1 and aflatoxin B1[J]. Environmental Chemistry, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
Citation: SUN Qing, LI Fuxue, JIA Yujun, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of an ELISA kit for simultaneous determination of aflatoxin M1 and aflatoxin B1[J]. Environmental Chemistry, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
shu

黄曲霉毒素M1和黄曲霉毒素B1总量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），国家自然科学基金（21576023），中央高校基本科研业务费专项资金（FRF-AS-15-005）资助.

Development and application of an ELISA kit for simultaneous determination of aflatoxin M1 and 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: Supported by the National High Technology Research and Development Program of China (2012AA101609-5), National Natural Science Foundation of China(21576023)and Fundamental Research Funds for the Central Universities(FRF-AS-15-005).

  • 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·mL-1，IC50为211 pg·mL-1，线性范围为50-1500 pg·mL-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.
    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·mL-1 and 211 pg·mL-1, respectively. The linear range was between 50 pg·mL-1 and 1500 pg·mL-1 for AFM1, Cross reactivities for AFM1 and AFB1 were 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.
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  • 收稿日期:  2016-04-14
  • 刊出日期:  2016-12-15
孙清, 李馥雪, 贾玉军, 邓乾民, 刘杰民, 时国庆. 黄曲霉毒素M1和黄曲霉毒素B1总量酶联免疫试剂盒的研制及应用[J]. 环境化学, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
引用本文: 孙清, 李馥雪, 贾玉军, 邓乾民, 刘杰民, 时国庆. 黄曲霉毒素M1和黄曲霉毒素B1总量酶联免疫试剂盒的研制及应用[J]. 环境化学, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
shu
SUN Qing, LI Fuxue, JIA Yujun, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of an ELISA kit for simultaneous determination of aflatoxin M1 and aflatoxin B1[J]. Environmental Chemistry, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
Citation: SUN Qing, LI Fuxue, JIA Yujun, DENG Qianmin, LIU Jiemin, SHI Guoqing. Development and application of an ELISA kit for simultaneous determination of aflatoxin M1 and aflatoxin B1[J]. Environmental Chemistry, 2016, 35(12): 2501-2510. doi: 10.7524/j.issn.0254-6108.2016.12.2016041401
shu

黄曲霉毒素M1和黄曲霉毒素B1总量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:  2016-04-14
基金项目:

国家高技术研究发展计划（2012AA101609-5），国家自然科学基金（21576023），中央高校基本科研业务费专项资金（FRF-AS-15-005）资助.

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·mL-1，IC50为211 pg·mL-1，线性范围为50-1500 pg·mL-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.

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