基于单克隆抗体的间接竞争酶联免疫吸附测定法用于检测食用动物组织和饲料中的硝基咪唑类药物。
Development of a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for nitroimidazoles in edible animal tissues and feeds.
作者信息
Han Wei, Pan Yuanhu, Wang Yulian, Chen Dongmei, Liu Zhenli, Zhou Qi, Feng Liang, Peng Dapeng, Yuan Zonghui
机构信息
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
出版信息
J Pharm Biomed Anal. 2016 Feb 20;120:84-91. doi: 10.1016/j.jpba.2015.12.017. Epub 2015 Dec 15.
The misuse of nitroimidazoles (NDZs) can lead to NDZs residues in edible animal tissues, which would be harmful to consumer health. To quickly monitor NDZs residues in edible animal tissues and feed, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) with a simple sample preparation method and clean-up was developed in the present study. At first, a broad-specificity monoclonal antibody, 1D5, against NDZs has been produced, which the IC50 values of the NDZs, dimetridazole, ipronidazole, ronidazole hydroxydimetridazole, and hydroxyipronidazole, were 4.79μgL(-1), 0.47μgL(-1), 5.97μgL(-1), 23.48μgL(-1), and 15.03μgL(-1), respectively. The limit of detection of the method for the NDZ matrix calibration ranged from 4.2μgkg(-1) to 50.3μgkg(-1) in the feed matrices and from 0.11μgkg(-1) to 4.11μgkg(-1) in the edible animal tissues matrices. The recoveries of the NDZs were in the range of 75.5-111.8%. The CVs were less than 14.4%. A good correlation (r=0.9905) between the ELISA and HPLC-MS results of the tissues demonstrated the reliability of the developed ic-ELISA, which makes it a useful tool for screening of NDZs in animal edible tissue and feed.
硝基咪唑类药物（NDZs）的滥用会导致其在可食用动物组织中残留，这对消费者健康有害。为了快速监测可食用动物组织和饲料中的NDZs残留，本研究开发了一种基于单克隆抗体的间接竞争酶联免疫吸附测定法（ic-ELISA），该方法具有简单的样品制备方法和净化步骤。首先，制备了一种针对NDZs的广谱特异性单克隆抗体1D5，其中NDZs、二甲硝唑、异丙硝唑、罗硝唑、羟基二甲硝唑和羟基异丙硝唑的半数抑制浓度（IC50）值分别为4.79μg/L、0.47μg/L、5.97μg/L、23.48μg/L和15.03μg/L。该方法对NDZ基质校准的检测限在饲料基质中为4.2μg/kg至50.3μg/kg，在可食用动物组织基质中为0.11μg/kg至4.11μg/kg。NDZs的回收率在75.5%-111.8%之间。变异系数小于14.4%。组织的ELISA和HPLC-MS结果之间具有良好的相关性（r=0.9905），证明了所开发的ic-ELISA的可靠性，这使其成为筛选动物可食用组织和饲料中NDZs的有用工具。
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