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重金属汞酶联免疫检测方法的建立[J]. 食品工业科技, 2012, (16): 86-88. DOI: 10.13386/j.issn1002-0306.2012.16.050
引用本文: 重金属汞酶联免疫检测方法的建立[J]. 食品工业科技, 2012, (16): 86-88. DOI: 10.13386/j.issn1002-0306.2012.16.050
Development of enzyme-linked immunosorbent assay for the detection of Hg (II)[J]. Science and Technology of Food Industry, 2012, (16): 86-88. DOI: 10.13386/j.issn1002-0306.2012.16.050
Citation: Development of enzyme-linked immunosorbent assay for the detection of Hg (II)[J]. Science and Technology of Food Industry, 2012, (16): 86-88. DOI: 10.13386/j.issn1002-0306.2012.16.050

重金属汞酶联免疫检测方法的建立

Development of enzyme-linked immunosorbent assay for the detection of Hg (II)

  • 摘要: 建立了重金属汞离子的间接竞争酶联免疫(IC-ELISA)分析方法。由于汞离子不具有免疫原性,因此以异硫氰酸苄基乙二胺四乙酸(ITCBE)为双功能螯合剂连接汞离子和钥孔血蓝蛋白(KLH)作为免疫原,免疫新西兰大白兔获得特异性抗体。用获得的抗体建立汞的ELISA分析方法。检测限为0.45μg/L,灵敏度为4.10μg/L。特异性实验表明,抗体对镍和铜的交叉反应率分别为10.8%和13.5%,与其他金属均无明显交叉反应。本实验以白芷、胡萝卜和皮皮虾为基质,添加回收实验表明,白芷、胡萝卜、皮皮虾回收率均在70%~120%。使用电感耦合等离子体质谱(ICP-MS)验证方法的准确性,仪器检测结果和建立的ELISA分析方法具有很好的相关性(相关系数为0.988),表明建立的方法具有很好的准确性。 

     

    Abstract: An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed for the rapid detection of Hg (II) . As the mercury ion was not immunogenic, so mercury was linked to the keyhole limpet hemocyanin (KLH) with 1- (4-isothiocyanobenzyl) ethylenediamine -N, N, N', N'-tetraacetic acid (ITCBE) as immunogen. For the developed ELISA, the limit of detection and sensitivity were 0.45μg/L and 4.10μg/L, respectively. The cross-reactivity against other metals were all low, except for copper and nickel, which had10.8%, 13.5% corss-reactivity, respectively. The recoveries of dahurica, carrot and Pipi shrimp sample were ranging from 70% to 120%. The accuracy of the developed IC-ELISA method was validated by inductively coupled plasma mass spectrometry (ICP-MS) . The developed ELISA for dahurica sample showed high correlation with ICP-MS (the correlation coefficients were 0.988) . Thus, it was demonstrated that the established immunoassay for the detection of Hg (II) was reliable.

     

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