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微珠芯片免疫法测定食品中氯霉素残留
Quantitation and Fast Immunoassay of Chloramphenicol Residues in Foodstuff by Microbeads Array
【摘要】 采用微珠芯片免疫分析法快速定量检测食品中的氯霉素残留。玻璃微珠经过表面化学修饰后形成氨基化表面,在碳二亚胺(EDC)催化作用下固定琥珀酸氯霉素,随后加入氯霉素单克隆抗体和氯霉素标准品或待测样品的混合物,待抗体抗原完全反应后加入荧光标记二抗示踪。绘制氯霉素浓度与荧光信号强度的标准曲线,然后根据标准曲线对待测样品中氯霉素进行定量分析。方法检测限为0.01μg.L-1;在0.05μg.L-1及0.2μg.L-1两个添加水平,加标回收率分别为92%~110%和92.5~108.5%;重复性实验CV分别为6.33%和5.99%;重现性实验CV分别为10.11%和9.12%。该方法具有良好的灵敏度、准确度和精密度,符合快速、高通量检测氯霉素残留的要求,并为实现介观流控免疫分析提供了基础。
【Abstract】 A microbeads array method for the quantitation and fast immunoassay of chloramphenicol(CAP) residues in foodstuff has been established.Catalyzed by EDC,chloramphenicol succinate is covalent combined on the amination surfaces of microbeads formed through chemical modification.After immobilization,the mixture of antibody for chloramphenicol and the standard or sample are added.Then the Cy5 labeled secondary antibody is added for tracing after antigen-antibody reaction.The calibration curve of chloramphenicol concentration and fluorescence intensity is established.With the calibration curve,the CAP concentration of the unknown sample can be guantitatively detected.The limit of detection(LOD)is 0.01 μg·L-1;On the spiked levels of 0.05 μg·L-1 and 0.2 μg·L-1,the recoveries are 92%~110% and 92.5%~108.5%,with the CV for the repeatability of 6.33% and 5.99% and the CV for reproducibility of 10.11% and 9.12%.The method is suitable for the rapid detection of chloramphenicol residues in foodstuff.
- 【文献出处】 化学与生物工程 ,Chemistry & Bioengineering , 编辑部邮箱 ,2008年09期
- 【分类号】TS207.53
- 【被引频次】7
- 【下载频次】169