制备单克隆抗体并建立酶联免疫吸附测定法检测动物源性食品中的头孢噻呋。
Preparation of monoclonal antibody and development of an enzyme-linked immunosorbent assay for the detection of ceftiofur in animal-derived foods.
机构信息
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues in Foods, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, China.
出版信息
J Pharm Biomed Anal. 2021 Nov 30;206:114378. doi: 10.1016/j.jpba.2021.114378. Epub 2021 Sep 17.
Ceftiofur (CEF) residues in animal-derived foods are of great concern to farmers, regulatory agencies and consumers. In this study, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) method was established to quickly monitor CEF residues in edible animal tissues using an easy sample preparation procedure. A monoclonal antibody, 4D5, against CEF has been produced at first, which had IC values for CEF, ceftriaxone, cefquinome, cefotaxime and desfuroylceftiofur of 0.78 μg/L, 0.73 μg/L, 13.6 μg/L, 8.99 μg/L and 8.89 μg/L, respectively. The limit of detection (LOD) and limit of quantitation (LOQ) in artificially contaminated animal-derived foods were 0.12-0.19 μg/L and 0.20-0.30 μg/L. The recovery rates were in the range of 89.7-109.0%. The CVs were less than 6.7%. A good correlation (R= 0.9994) between the ic-ELISA and UPLC-MS/MS showed the reliability of the developed ic-ELISA. The ic-ELISA produces a sensitive, accurate and low-cost tool for the screening of residues of CEF in animal-derived foods.
动物源性食品中的头孢噻呋（CEF）残留问题引起了农民、监管机构和消费者的高度关注。本研究采用一种简单的样品制备程序，建立了一种间接竞争酶联免疫吸附测定法（ic-ELISA），用于快速监测食用动物组织中的 CEF 残留。首先制备出了针对 CEF 的单克隆抗体 4D5，其对 CEF、头孢曲松、头孢喹肟、头孢噻肟和去呋喃甲酰头孢噻呋的 IC 值分别为 0.78μg/L、0.73μg/L、13.6μg/L、8.99μg/L 和 8.89μg/L。在人工污染的动物源性食品中，该方法的检测限（LOD）和定量限（LOQ）分别为 0.12-0.19μg/L 和 0.20-0.30μg/L。回收率在 89.7-109.0%范围内。CV 小于 6.7%。ic-ELISA 与 UPLC-MS/MS 的相关性良好（R=0.9994），表明所建立的 ic-ELISA 具有可靠性。ic-ELISA 为动物源性食品中 CEF 残留的筛选提供了一种灵敏、准确且低成本的工具。
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