Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University , Beijing, PR China.
Products Development Department, Beijing WDWK Biotechnology Co. Ltd , Beijing, PR China.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2020 Oct;37(10):1633-1639. doi: 10.1080/19440049.2020.1787527. Epub 2020 Jul 28.
Diclazuril, a broad-spectrum anticoccidial drug, may be accumulated in edible tissues of animals through illegal use, which poses potential threats to human health through the food chain. In this study, an innovative hapten was designed and an immunogen of diclazuril was successfully synthesised with keyhole limpet haemocyanin as carrier protein; then a monoclonal antibody with high specificity was obtained. Furthermore, based on the novel antibody, a one-step indirect competitive enzyme-linked immunosorbent assay (icELISA) was established for rapid and specific detection of diclazuril residues. Compared with the traditional icELISA method, this method saves at least 0.5 hours and one washing step. Under the optimal conditions, the one-step icELISA for diclazuril exhibited good performance with a 50% inhibition concentration (IC) value of 0.952 μg/kg. The average recoveries of the icELISA ranged from 73.1% to 115.5% with the coefficient of variation lower than 12.7%, which was evaluated by detecting spiked animal-origin food samples. Finally, the one-step icELISA shows a good correlation with an ultra-high liquid chromatography-tandem mass spectrometry method. Those results demonstrate that the one-step icELISA developed for diclazuril detection is time-saving, low-cost, specific, sensitive, and reliable. It shows good potential for social, environmental, and economic benefits in future use.
地克珠利是一种广谱抗球虫药物，由于非法使用，可能会在动物可食用组织中蓄积，通过食物链对人类健康造成潜在威胁。本研究设计了一种新的半抗原，并以钥孔贻贝血红蛋白为载体蛋白成功合成了地克珠利免疫原；然后获得了一种具有高特异性的单克隆抗体。此外，基于该新型抗体，建立了一种用于快速、特异性检测地克珠利残留的一步间接竞争酶联免疫吸附测定法（icELISA）。与传统的 icELISA 方法相比，该方法至少节省了 0.5 小时和一个洗涤步骤。在最佳条件下，一步 icELISA 法检测地克珠利的 50%抑制浓度（IC）值为 0.952μg/kg。icELISA 法检测动物源性食品样品的平均回收率为 73.1%至 115.5%，变异系数低于 12.7%。最后，一步 icELISA 法与超高效液相色谱-串联质谱法具有良好的相关性。这些结果表明，开发的一步 icELISA 法检测地克珠利具有省时、低成本、特异性强、灵敏度高、可靠性好的特点。在未来的应用中具有良好的社会效益、环境效益和经济效益潜力。
