A Helper Antibody-Based Competitive Fluorescence Immunochromatographic Assay for Quantitative Detection of Florfenicol in Poultry Eggs.

IF 1.7 4区 农林科学 Q3 CHEMISTRY, ANALYTICAL
Enhui Zhang, Bochao Liu, Jinhui Lu, Chaolan Liang, Fang Zhao, Jinfeng Li, Tingting Li, Chengyao Li, Ling Zhang
{"title":"A Helper Antibody-Based Competitive Fluorescence Immunochromatographic Assay for Quantitative Detection of Florfenicol in Poultry Eggs.","authors":"Enhui Zhang,&nbsp;Bochao Liu,&nbsp;Jinhui Lu,&nbsp;Chaolan Liang,&nbsp;Fang Zhao,&nbsp;Jinfeng Li,&nbsp;Tingting Li,&nbsp;Chengyao Li,&nbsp;Ling Zhang","doi":"10.1093/jaoacint/qsad044","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Florfenicol (FF) is a chloramphenicol analogue used in animals, and florfenicol amine (FFA) is the main metabolite of FF. However, their residues in agricultural products are harmful to human health. A highly specific and sensitive assay for FF/FFA detection needs to be developed since the traditional detection methods are low in sensitivity.</p><p><strong>Objective: </strong>In this study, a new method for rapid quantification of FF/FFA in poultry eggs by helper antibody-based fluorescent immunochromatographic assay (HAFIA) was established.</p><p><strong>Methods: </strong>Triple antibodies including a primary monoclonal antibody (mAb) specific to the targets FF and FFA, a secondary polyclonal antibody (pAb) labeled with europium nanoparticles (EuNPs), and a helper monoclonal antibody (hAb), reacting with pAb but not with the mAb or the target antigen, are designed, which can form structural aggregation complexes in microwells with a single step of reactions. By loading the reaction sample solution, the triple-antibodies (mAb-pAb-hAb)-EuNPs complexes migrate to the test (T) line on the nitrocellulose membrane of testing strip and are competitively captured by the immobilized FF-bovine serum album (BSA) conjugates on the membrane and the FF/FFA targets in the sample solution.</p><p><strong>Results: </strong>Fluorescence on the T line is read by a portable fluorescent strip reader in 10 min, and the result is given as the ratio of fluorescent intensities on the T and control (C) lines. This new fluorescent testing strip, with amplified signal from the triple-antibody complex, has 50-fold higher sensitivity than conventional colloidal gold-lateral flow immunoassays (CG-LFIAs), and can detect as low as 0.01 ng/mL FF and 0.1 ng/mL FFA targets from egg samples.</p><p><strong>Conclusion: </strong>The developed competitive fluorescent immunochromatography method based on auxiliary antibodies has the advantages of high sensitivity and specificity for the rapid and quantitative detection of FF/FFA in poultry eggs.</p><p><strong>Highlights: </strong>Newly designed helper antibody and portable device were applied to quantitative detection. HAFIA tests egg samples and results can be obtained in 10 minutes. HAFIA has the advantages of being more convenient, faster and does not require professional laboratory personnel.</p>","PeriodicalId":15003,"journal":{"name":"Journal of AOAC International","volume":null,"pages":null},"PeriodicalIF":1.7000,"publicationDate":"2023-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of AOAC International","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1093/jaoacint/qsad044","RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0

Abstract

Background: Florfenicol (FF) is a chloramphenicol analogue used in animals, and florfenicol amine (FFA) is the main metabolite of FF. However, their residues in agricultural products are harmful to human health. A highly specific and sensitive assay for FF/FFA detection needs to be developed since the traditional detection methods are low in sensitivity.

Objective: In this study, a new method for rapid quantification of FF/FFA in poultry eggs by helper antibody-based fluorescent immunochromatographic assay (HAFIA) was established.

Methods: Triple antibodies including a primary monoclonal antibody (mAb) specific to the targets FF and FFA, a secondary polyclonal antibody (pAb) labeled with europium nanoparticles (EuNPs), and a helper monoclonal antibody (hAb), reacting with pAb but not with the mAb or the target antigen, are designed, which can form structural aggregation complexes in microwells with a single step of reactions. By loading the reaction sample solution, the triple-antibodies (mAb-pAb-hAb)-EuNPs complexes migrate to the test (T) line on the nitrocellulose membrane of testing strip and are competitively captured by the immobilized FF-bovine serum album (BSA) conjugates on the membrane and the FF/FFA targets in the sample solution.

Results: Fluorescence on the T line is read by a portable fluorescent strip reader in 10 min, and the result is given as the ratio of fluorescent intensities on the T and control (C) lines. This new fluorescent testing strip, with amplified signal from the triple-antibody complex, has 50-fold higher sensitivity than conventional colloidal gold-lateral flow immunoassays (CG-LFIAs), and can detect as low as 0.01 ng/mL FF and 0.1 ng/mL FFA targets from egg samples.

Conclusion: The developed competitive fluorescent immunochromatography method based on auxiliary antibodies has the advantages of high sensitivity and specificity for the rapid and quantitative detection of FF/FFA in poultry eggs.

Highlights: Newly designed helper antibody and portable device were applied to quantitative detection. HAFIA tests egg samples and results can be obtained in 10 minutes. HAFIA has the advantages of being more convenient, faster and does not require professional laboratory personnel.

基于辅助抗体的竞争荧光免疫层析法定量检测禽蛋中氟苯尼考。
背景:氟苯尼考(FF)是一种用于动物的氯霉素类似物,氟苯尼考胺(FFA)是FF的主要代谢物。然而,它们在农产品中的残留对人体健康有害。由于传统的检测方法灵敏度较低,需要开发一种高特异性和高灵敏度的FF/FFA检测方法。目的:建立基于辅助抗体的荧光免疫层析法(HAFIA)快速定量测定禽蛋中FF/FFA的新方法。方法:设计靶向FF和FFA的一级单克隆抗体(mAb)、铕纳米颗粒标记的二级多克隆抗体(pAb)和辅助单克隆抗体(hAb),与pAb反应而不与mAb或靶抗原反应,通过一步反应在微孔中形成结构聚集复合物。通过加载反应样品溶液,三抗体(mab - ab - hab)-EuNPs复合物迁移到测试条上的硝化纤维素膜上的测试(T)线上,并被固定在膜上的FF-bovine serum album (BSA)偶联物和样品溶液中的FF/FFA靶点竞争性捕获。结果:便携式荧光条带读取器在10min内读取T线荧光,结果为T线与对照(C)线荧光强度之比。这种新的荧光测试条具有来自三抗体复合物的放大信号,灵敏度比传统的胶体金-侧流免疫测定法(CG-LFIAs)高50倍,并且可以从鸡蛋样品中检测低至0.01 ng/mL的FF和0.1 ng/mL的FFA靶标。结论:建立的以辅助抗体为基础的竞争荧光免疫层析法具有灵敏度和特异性高的优点,可快速定量检测禽蛋中FF/FFA。亮点:采用新设计的辅助抗体和便携式仪器进行定量检测。HAFIA测试鸡蛋样本,结果可以在10分钟内得到。HAFIA具有更方便、快捷、不需要专业实验室人员的优点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Journal of AOAC International
Journal of AOAC International 医学-分析化学
CiteScore
3.10
自引率
12.50%
发文量
144
审稿时长
2.7 months
期刊介绍: The Journal of AOAC INTERNATIONAL publishes the latest in basic and applied research in analytical sciences related to foods, drugs, agriculture, the environment, and more. The Journal is the method researchers'' forum for exchanging information and keeping informed of new technology and techniques pertinent to regulatory agencies and regulated industries.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信