Wang Zhaopeng, Yu Xuezhi, Ma Licai, Liu Hebing, Ding Shuangyang, Wang Zhanhui, Zhang Xiya, Shen Jianzhong, Wen Kai
a College of Veterinary Medicine, China Agricultural University , Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety , Beijing , People's Republic of China.
b Research and Development Department, Beijing WDWK Biotechnology Company, Ltd ., Beijing , People's Republic of China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2018 Jul;35(7):1247-1256. doi: 10.1080/19440049.2018.1447693. Epub 2018 May 23.
In this work, high affinity polyclonal antibodies for ribavirin (RBV) from new haptens were prepared and were used to analyse RBV residues in chicken muscle, eggs and duck muscle. The new haptens were synthesised with different spacers, and the best antibody was obtained with an IC value as low as 0.61 ng/mL in indirect competitive enzyme-linked immunosorbent assay (ELISA). The cross-reactivities with another five antiviral drugs including amantadine, rimantadine, moroxydine, zanamivir and oseltamivir were less than 0.1%, which indicated the good specificity of the antibody. An ELISA was developed based on the antibody and applied to detect RBV in multi-food matrices. The sample preparation prior to detection only needed simple dilution after trichloroacetic acid extraction. The limits of detection were 1.07, 1.18 and 1.03 μg/kg in chicken muscle, eggs and duck muscle, respectively. Recoveries ranged from 89.0% to 112.7% with coefficients of variation below 13.0%. Ten blind samples of chicken muscle were analysed simultaneously by ELISA and liquid chromatography-tandem mass spectrometry, and a good correlation between the methods was observed. The results indicated that the high affinity antibody could be applied for the simple and fast detection of RBV in multi-food matrices.
在本研究中，制备了针对利巴韦林（RBV）的新型半抗原高亲和力多克隆抗体，并用于分析鸡肉、鸡蛋和鸭肉中的RBV残留量。新型半抗原通过不同间隔臂合成，在间接竞争酶联免疫吸附测定（ELISA）中获得了IC值低至0.61 ng/mL的最佳抗体。该抗体与金刚烷胺、金刚乙胺、吗啉胍、扎那米韦和奥司他韦等另外五种抗病毒药物的交叉反应率均低于0.1%，表明该抗体具有良好的特异性。基于该抗体开发了一种ELISA方法，并应用于多种食品基质中RBV的检测。检测前的样品制备仅需在三氯乙酸提取后进行简单稀释。鸡肉、鸡蛋和鸭肉中的检测限分别为1.07、1.18和1.03 μg/kg。回收率在89.0%至112.7%之间，变异系数低于13.0%。同时采用ELISA和液相色谱-串联质谱法对10份鸡肉盲样进行分析，两种方法之间具有良好的相关性。结果表明，该高亲和力抗体可用于多种食品基质中RBV的简单快速检测。
