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基于高选择性单克隆抗体的侧向流动免疫分析法用于可视化和灵敏测定蔬菜中的唑类杀菌剂丙环唑。

Highly selective monoclonal antibody-based lateral flow immunoassay for visual and sensitive determination of conazole fungicides propiconazole in vegetables.

作者信息

Xu Ze-Ke, Meng Jing-Nan, Lei Yi, Yang Xing-Xing, Yan Yi-Yong, Liu Hai-Hong, Lei Hong-Tao, Wang Ting-Cai, Shen Xing, Xu Zhen-Lin

机构信息

Guangdong Provincial Key Laboratory of Food Quality and Safety/ Guangdong Laboratory of Lingnan Modern Agriculture, South China Agricultural University, Guangzhou, China.

Department of Organic Substances Analyisis, Guangdong Institute of Food Inspection, Guangzhou, China.

出版信息

Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2022 Jan;39(1):92-104. doi: 10.1080/19440049.2021.1976423. Epub 2021 Oct 26.

Abstract

The conazole fungicide propiconazole is frequently found in vegetables although usage is not allowed. To overcome the high-cost and time-consuming labour requirements of instrumental methods, we developed a simple and visual lateral flow immunoassay for the sensitive determination of propiconazole. A hapten was carefully designed to raise a monoclonal antibody against propiconazole. Bal b/c mice were immunised with the hapten-carrier protein conjugate and a specific monoclonal antibody (mAb) was produced. Based on this mAb, a sensitive immunochromatographic strip assay (ICA) was established for rapid screening of propiconazole in vegetable samples. After optimisation of analytical parameters, the ICA strip showed a detection limit of 0.13 ng g and a linear range from 0.5 to 80 ng g using a strip reader. The assay also can be read by the naked eye with a visual limit of detection of 80 ng g. The recoveries for spiked vegetable samples by ICA ranged from 85.2% to 114.9%, with a coefficient of variation less than 11.7%. The assay time is within 45 min for a single sample including the sample pre-treatment. For spiked and blind samples, the detection capability of ICA was equivalent to liquid chromatography-mass spectrometry.

摘要

尽管不允许使用,但蔬菜中经常能检测到唑类杀菌剂丙环唑。为克服仪器分析方法成本高且耗时的问题,我们开发了一种简单直观的侧向流动免疫分析法,用于灵敏检测丙环唑。精心设计了一种半抗原,以产生抗丙环唑的单克隆抗体。用半抗原 - 载体蛋白偶联物免疫Bal b/c小鼠,制备出特异性单克隆抗体(mAb)。基于该单克隆抗体,建立了一种灵敏的免疫层析试纸条检测法(ICA),用于快速筛查蔬菜样品中的丙环唑。优化分析参数后,使用试纸条读数仪时,ICA试纸条的检测限为0.13 ng/g,线性范围为0.5至80 ng/g。该检测法也可用肉眼读取,目视检测限为80 ng/g。ICA对加标蔬菜样品的回收率为85.2%至114.9%变异系数小于11.7%。包括样品预处理在内,单个样品的检测时间在45分钟以内。对于加标样品和盲样,ICA的检测能力与液相色谱 - 质谱法相当。

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