Development of Indirect Competitive ELISA and Visualized Multicolor ELISA Based on Gold Nanorods Growth for the Determination of Zearalenone.

Tianyu Ma,Xiao Yang,Kaixin Liu,Mingfei Pan,Shuo Wang,Jingying Yang

doi:10.3390/foods10112654

Tianyu Ma, Xiao Yang + Show 4 more

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https://doi.org/10.3390/foods10112654

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Abstract

In this study, a zearalenone (ZEN) hapten was designed and prepared against the mycotoxin ZEN, and the original coating ZEN-ovalbumin (ZEN-OVA) was prepared by conjugation with OVA. Based on the gold nanorods (AuNRs) of uniform size and stable properties synthesized by the seed-mediated method, the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and the AuNRs growth-based multicolor ELISA for detecting ZEN toxin were further established. Under the optimal experimental conditions, the coating amount of ZEN-OVA: 0.025 μg/well, antibody (Ab) dilution factor: 32,000 times, blocking solution: 0.5% skimmed milk powder, enzyme-labeled secondary Ab diluted 10,000 times, and a pH of the PBS buffer at 7.4, the sensitivity (IC50) of the established ic-ELISA for ZEN detection reached 0.85 ± 0.04 μg/L, and the limit of detection (IC15) reached 0.22 ± 0.08 μg/L. In the multicolor ELISA based on the growth of AuNRs, as the content of ZEN increased, the mixed solution exhibited a significant color change from brownish red to colorless. ZEN concentration as low as 0.1 μg/L can be detected by the naked eye (brown red to dark gray). This study provided an effective analysis strategy for the rapid screening and accurate monitoring of the ZEN contaminant in foods.

Highlights

Zearalenone (ZEN) is a nonsterol estrogen mycotoxin mainly produced by Fusarium genera, which is widely found in grain crops, such as corn, wheat, and barley [1,2]
It has been reported that the potential etiological mechanism of breast cancer involves changes in the cytochrome P450 (CYP) enzyme, which is related to ZEN [7]
Ltd. (Shandong, China). 3,3,5,5-tetramethylbenzidine (TMB), horseradish peroxidase (HRP) goat anti-mouse IgG conjugated (1.0 mg/mL), alkaline phosphatase (AP) goat anti-mouse IgG conjugated (3.5 mg/mL), aflatoxin B1 (AFB1), ZEN and the structural analogues (α-zearalenol, αzearalanol, β-zearalenol, zearalanone, β-zearalanol) (1.0 mg/L) were purchased from Sigma-Aldrich

Summary

Introduction

Zearalenone (ZEN) is a nonsterol estrogen mycotoxin mainly produced by Fusarium genera, which is widely found in grain crops, such as corn, wheat, and barley [1,2]. ZEN can cause excessive estrogen syndrome in animals such as pig, poultry, and humans, as well as immunotoxicity, genotoxicity, and suspected carcinogenicity [5,6]. Yu et al have demonstrated that the ZEN on 2,3,7,8-Tetrachlorodibenzop-dioxin (TCDD)-induced CYP1A1 activity and gene expression involved the estrogen receptor pathway [8]. Due to its similar structure to endogenous estrogen, ZEN can show estrogen activity in vivo and competitively bind with estrogen receptors, affecting estrogen secretion in humans or animals, resulting in reproductive organ abnormalities, infertility, abortion, and other diseases [9,10,11]. The development of effective strategies for ZEN detection in foods is of great significance for protecting the health of humans and animals

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