鸡肉中氟苯尼考ELISA检测方法的建立及应用

被引：6

作者：

孙法良

刁有祥

孙宁

马艳芳

纪巍

机构：

[1] 山东农业大学动物科技学院

来源：

中国农业科学 | 2009年 / 05期

关键词：

氟苯尼考; 残留检测; 间接竞争ELISA;

D O I：

暂无

中图分类号：

S859.84 [动物性食品中化学物残留];

学科分类号：

0906 ;

摘要：

【目的】制备抗氟苯尼考(FFC)的高亲和力特异性抗体,建立检测氟苯尼考的间接竞争ELISA新方法。【方法】氟苯尼考分别与牛血清白蛋白(BSA)和人血清白蛋白(HSA)经混合酸酐法偶联,得到免疫抗原和包被抗原,其偶联比分别为14.4和14.7。用免疫原FFC-HS-BSA免疫新西兰大白兔获得了效价较高的特异性抗体。建立并优化了间接竞争ELISA检测方法。【结果】抗血清效价为1:102400。最佳包被抗原浓度为0.5μg·ml-1,最佳抗体工作浓度为1:6400,酶标二抗的工作浓度为1:20000。回归方程Y=-0.1516X+0.788(R2=0.9859),检测范围为0.18～500ng·ml-1,检出限为0.18ng·ml-1,批内和批间平均变异系数分别为4.86%和7.77%。交叉反应试验中,抗血清与FFC结构相似的氯霉素和甲砜霉素交叉反应率分别为0.094%和0.098%,与其它药物交叉反应率均小于0.01%,表明该方法具有很强的特异性。FFC以浓度0.18～500ng·g-1在鸡肉中添加,回收率为84.14%～106.1%,变异系数为2.1%～5.6%。【结论】成功获得了高效价、高特异性的抗FFC抗体,建立了鸡肉中氟苯尼考残留检测的ELISA方法。结果表明,所建立的检测方法具有灵敏、准确、简便、快速的特点。

引用

页码：1813 / 1819

页数：7

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