Heterologous-coating antigen enhancing the sensitivity of enzyme-linked immunosorbent assay for detection of mebendazole residues

Abstract

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 (IC₂₀–IC₈₀) 0.34–10.54 ng/mL, an IC₅₀ 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 (IC₅₀ 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.

Keywords:

• Heterologous coating antigen
• rabbit IgG
• chicken IgY
• mebendazole
• amino-mebendazole

Ethical approval

The protocol for carrying out animal experiments was approved by the Ethics Committee of Shanxi Agricultural University for the use of laboratory Animals, and all the ethical requirements to conduct the experiment were met.

Author contributions

Shengrui Shi and Jinxin He: Conceptualization and validation. Shengrui Shi and Fujun Yang: Data curation and formal analysis. Xiaorong Chen and Shengrui Shi: Software and Methodology. Yayan Yang and Fujun Yang: Visualization and Software. Jinxin He and Shaopeng Gu: Writing and Editing.

Disclosure statement

All the authors declare that they have no conflict of interest.

Data availability statement

The data that support the findings of this study are available from the corresponding author by request.

Additional information

Funding

This work was supported by the Project of Science and Technology Innovation Fund of Shanxi Agricultural University [2020BQ45]. Innovation and Entrepreneurship Training Project for College Students in Shanxi Province [S202110113143]. Innovation Projects of College of Veterinary Medicine, Shanxi Agricultural University [DYXY2021DC08; DY-Q012]. Fund for Shanxi “1331 Project” [20211331-13]. Project of Scientific Research for Excellent Doctors [SXYBKY2019026], Shanxi Province, China.