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建立一种高灵敏度的基于单克隆抗体的间接竞争酶联免疫吸附分析法,用于饲料中维吉霉素的检测。

Development of a highly sensitive monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for the detection of avilamycin in feed.

机构信息

College of Food Science and Biotechnology, Zhejiang Gongshang University, Zhejiang, P. R. China.

出版信息

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2022 Mar;39(3):488-498. doi: 10.1080/19440049.2021.2017003. Epub 2022 Jan 21.

Abstract

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed using a highly sensitive and specific monoclonal antibody against avilamycin. The immunising antigen synthesised through the carbonyldiimidazole (CDI) method was used to prepare antibodies. The established ic-ELISA, after optimisation of conditions, possessed a half maximum inhibitory concentration (IC) value of 7.44 ng mL-1 and the detection limit of 0.21 ng mL in the standard curve. The spiked experiments indicated that the limits of detection were 1.86 µg kg and 2.31 µg kg in swine feed and chicken feed, respectively. In addition, the average recoveries ranged from 74.7 to 105.4% with the coefficient of variation less than 11%. The good correlation (R = 0.9818) between the result of ic-ELISA and HPLC demonstrated the reliability of the developed ic-ELISA, which showed that the ic-ELISA developed here achieves the aim of strengthening the monitoring of avilamycin residues and the inspection of import and export related products.

摘要

建立了一种基于抗维吉霉素单克隆抗体的间接竞争酶联免疫吸附分析方法(ic-ELISA)。该方法使用碳化二亚胺(CDI)法合成的免疫原制备抗体。优化条件后,建立的 ic-ELISA 的半最大抑制浓度(IC)值为 7.44ng/mL,标准曲线的检测限为 0.21ng/mL。加标实验表明,在猪饲料和鸡饲料中的检测限分别为 1.86μg/kg 和 2.31μg/kg。此外,平均回收率在 74.7%至 105.4%之间,变异系数小于 11%。ic-ELISA 与 HPLC 结果的良好相关性(R=0.9818)表明该方法可靠,表明本研究建立的 ic-ELISA 达到了加强维吉霉素残留监测和进出口相关产品检测的目的。

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