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牛奶中喹诺酮类药物残留的AlphaLISA检测方法的研究

马立才 邢维维 于莹 贾良曦 秦誉 金兴 丁亚芳

食品工业科技2024,Vol.45Issue(14):264-270,7.
食品工业科技2024,Vol.45Issue(14):264-270,7.DOI:10.13386/j.issn1002-0306.2023080295

牛奶中喹诺酮类药物残留的AlphaLISA检测方法的研究

AlphaLISA Detection Method for Quinolone Residues in Milk

马立才 1邢维维 1于莹 1贾良曦 1秦誉 1金兴 1丁亚芳1

作者信息

  • 1. 北京维德维康生物技术有限公司,北京 100095
  • 折叠

摘要

Abstract

This article aimed to establish an amplified luminescent proximity homogeneous assay linked immune sorbent assay(AlphaLISA)for detecting quinolones(QNS)residues in milk.The EDC/NHS active ester method was used to connect 6-aminocaproic acid as the spacer arm on the norfloxacin molecule to prepare biotin-modified norfloxacin hapten(Biotin-6AS-NOR),and QNS monoclonal antibody was coupled on the receptor microsphere.Biotin-6AS-NOR could compete with QNS in the sample to bind quinolone antibodies.A competitive indirect AlphaLISA for QNS residues in milk was established by optimizing the parameters including QNS antibody coupling amount on receptor microspheres,Biotin-6AS-NOR,the dosage of donor/receptor microspheres,and dilution schemes for milk samples.By using the 1∶5 milk sample dilution scheme,the limit of detection and the limit of quantification of the established AlphaLISA for QNS in milk were 0.19 ng/mL and 0.46 ng/mL,respectively.The recovery values ranged from 85.97%and 110.11%,and the intraday and interday variation were both less than 10%.The cross-reactivity values for norfloxacin,enrofloxacin,ciprofloxacin,ofloxacin,pefloxacin,ofloxacin,and lomefloxacin were all higher than 70%.In conclusion,the established AlphaLISA showed high sensitivity,accuracy and good repeatability,without the need of tedious wash steps throughout the detection process;therefore,it is easy to achieve automated detection of chemical hazards residue in milk and has high application and promotion value.

关键词

喹诺酮类/半抗原/生物素/AlphaLISA

Key words

quinolones/hapten/biotin/AlphaLISA

分类

轻工纺织

引用本文复制引用

马立才,邢维维,于莹,贾良曦,秦誉,金兴,丁亚芳..牛奶中喹诺酮类药物残留的AlphaLISA检测方法的研究[J].食品工业科技,2024,45(14):264-270,7.

基金项目

石家庄市科技计划:生牛乳中危害因子高通量检测技术的研发与应用(221170243A). (221170243A)

食品工业科技

OA北大核心CSTPCD

1002-0306

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