磺胺二甲嘧啶间接竞争化学发光酶免疫分析法的建立及应用
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(1.广西壮族自治区兽医研究所,广西 南宁 530001; 2.南宁市动物卫生监督所,广西 南宁 530001; 3.广西壮族自治区兽药监察所,广西 南宁 530001)

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马玲 女 助理研究员 研究方向为分子免疫学与传染病学 E-mail:maling0120@sohu.com通讯作者:┣┣(中)通信作者┫┫吴健敏 女 研究员 研究方向为分子病毒学与动物传染病 E-mail:wu-jm20@163.com

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广西壮族自治区科技厅基本科研业务专项(桂科专项12-4);南宁市科学研究与技术开发计划-科技创新能力与条件建设项目(201102047B)


Development and application of indirect competitive chemiluminescence immunoassay for detecting sulfadimidine residue
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(Guangxi Veterinary Research Institute,Guangxi Nanning 530001,China)

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    摘要:

    建立一种更加灵敏的磺胺二甲嘧啶(SM2)免疫学检测方法。方法 本研究制备抗SM2单克隆抗体(SM2-mAb),用于建立SM2间接竞争化学发光酶免疫分析法(SM2-CLEIA),并检测主要动物食品中SM2残留。结果 本研究制备的SM2-mAb为IgG2b亚类,亲和常数为0.12×107 L/mol;SM2-CLEIA曲线在0.1~1 000 μg/L之间呈现良好的线性关系,相关系数r2=0.990 4,半数抑制浓度(IC50)为4.006 μg/L,检测限为0.174 μg/L,添加回收率为94.41%~104.40%,批内和批间变异系数分别为3.07%和8.22%,与磺胺脒等药物无交叉反应,与SM2-ELISA试剂盒的阴阳性符合率为100%。结论 建立了灵敏、特异、准确、检测范围宽的SM2-CLEIA检测方法。

    Abstract:

    To develop a more sensitive immunologic method for SM2 detection. MethodsAn indirect competitive chemiluminescence immunoassay was established with SM2 monoclonal antibody, and was used to detect the SM2 residue in animal derived foods. ResultsThe results indicated that the SM2-mAb belongs to subclass of IgG2b, and its affinity constant was 0.12×107L/mol. The detection limit of SM2-CLEIA was 0.174 μg/L with the linear range of 0.1 to 1 000 μg/L (r2=0.990 4), and IC50 was 4.006 μg/L. The recoveries were from 94.41% to 104.40%. The variability within and between batches were 3.07% and 8.22%, respectively. There was no significant cross reaction between SM2-mAb and SG or other drugs, and the results were 100% in accordance with those of the SM2-ELISA kit.ConclusionSM2-CLEIA was established, which was sensitive, specific, precise and with wide linear range.

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马玲,韦建兴,陆忠础,唐承明,李辉,李智红,张怡轩,甘浪帆,吴健敏.磺胺二甲嘧啶间接竞争化学发光酶免疫分析法的建立及应用[J].中国食品卫生杂志,2014,26(5):455-460.

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  • 收稿日期:2014-07-02
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  • 在线发布日期: 2014-10-16
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