College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi, PR China.
J Environ Sci Health B. 2022;57(11):883-889. doi: 10.1080/03601234.2022.2129938. Epub 2022 Oct 10.
The heterologous strategy could improve the sensitivity of competitive enzyme-linked immunosorbent assay (ELISA) for detection of chemical contaminants in food samples. In this study, the heterologous coating antigen ELISA was developed to evaluate its sensitivity for mebendazole (MBZ). Results showed that the heterologous ELISA had a linear range of (IC-IC) 0.34-10.54 ng/mL, an IC value of 1.83 ng/mL, and a limit of detection (LOD) of 0.13 ng/mL, in which the sensitivity of ELISA improved 1.7- and 2-fold (IC value dropping from 7.41 and 3.65 ng/mL to 4.27 and 1.83 ng/mL) than that of rabbit IgG- and chicken IgY-based homologous ELISA for MBZ, respectively. The heterologous coating antigen ELISA showed negligible cross reactivity (<0.2%) with its structural analogues, including hydroxy-MBZ, albendazole, oxfendazole, fenbendazole, and flubendazole, except the value of 72.6% for amino-MBZ. The average recoveries of MBZ spiked in pork and chicken muscle samples by the assay ranged from 83.7% to 109.8% and agreed well with those of high-performance liquid chromatography. The results suggested that using heterologous coating antigen could distinctly improve the sensitivity of ELISA for routine screening of MBZ residues in food samples.
异源策略可以提高竞争性酶联免疫吸附测定（ELISA）检测食品样品中化学污染物的灵敏度。在这项研究中，开发了异源包被抗原 ELISA 来评估其对苯硫咪唑（MBZ）的灵敏度。结果表明，异源 ELISA 的线性范围为（IC-IC）0.34-10.54ng/mL，IC 值为 1.83ng/mL，检测限（LOD）为 0.13ng/mL，其中 ELISA 的灵敏度比兔 IgG 和鸡 IgY 基于同源 ELISA 的灵敏度分别提高了 1.7 倍和 2 倍（IC 值从 7.41ng/mL 和 3.65ng/mL 降低到 4.27ng/mL 和 1.83ng/mL）。异源包被抗原 ELISA 与 MBZ 的结构类似物（包括羟基 MBZ、阿苯达唑、奥芬达唑、芬苯达唑和氟苯达唑）的交叉反应性可忽略不计（<0.2%），除了氨基 MBZ 的值为 72.6%。该测定法用于检测猪肉和鸡肉肌肉样品中 MBZ 加标回收率在 83.7%至 109.8%之间，与高效液相色谱法吻合良好。结果表明，使用异源包被抗原可以显著提高 ELISA 检测食品样品中 MBZ 残留的常规筛选灵敏度。
