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Sensitive fluorescence ELISA for the detection of zearalenone based on self-assembly DNA nanocomposites and copper nanoclusters

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Abstract

Zearalenone (ZEN), produced by Fusarium species, is a potential risk to human health. Traditional enzyme-linked immunosorbent assay (ELISA) is restricted due to low sensitivity for the detection of ZEN. Herein, enzyme nanocomposites (ALP-SA-Bio-ssDNA, ASBD) were prepared with the self-assembly strategy based on streptavidin-labeled alkaline phosphatase (SA-ALP) and dual-biotinylated ssDNA (B2-ssDNA). The enzyme nanocomposites improved the loading amount of ALP and catalyzed more ascorbic acid 2-phosphate to generate ascorbic acid (AA). Subsequently, Cu2+ could be reduced to copper nanoclusters (CuNCs) having strong fluorescence signal by AA with poly T. Benefiting from the high enzyme load of nanocomposites and the strong signal of CuNCs, the fluorescence ELISA was successfully established for the detection of ZEN. The proposed method exhibited lower limit of detection (0.26 ng mL−1) than traditional ELISA (1.55 ng mL−1). The recovery rates ranged from 92.00% to 108.38% (coefficient of variation < 9.50%) for the detection of zearalenone in corn and wheat samples. In addition, the proposed method exhibited no cross reaction with four other mycotoxins. This proposed method could be used in trace detection for food safety.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (82260644) and Natural Science Foundation of Jiangxi Province (20212BAB215041) and Pilot Demonstration Project of Ceilings in Funding for Provincial Science and Technology Program of Jiangxi Academy of Sciences (2021YSBG22023 and 2023YJC2006).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Resources, Nanchang University, 235, East Nanjing Road, Nanchang, 330047, China

    Xudong Jing, Sha Yu, Yanyan Tang, Jiaqi Yin, Juan Peng & Weihua Lai

  2. Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang, 330096, China

    Ganggang Zhang

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  1. Xudong Jing
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  2. Sha Yu
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  3. Ganggang Zhang
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  7. Weihua Lai
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Contributions

Xudong Jing: Conceptualization, Investigation, Software, Writing—original draft, Writing – review & editing. Sha Yu: Investigation, Software. Ganggang Zhang: Investigation, Software, Funding acquisition, Supervision, Writing – review & editing. Yanyan Tang: Investigation, Software. Jiaqi Yin: Investigation, Software. Juan Peng: Formal analysis, Methodology, Validation. Weihua Lai: Formal analysis, Methodology, Validation, Funding acquisition, Supervision, Writing – review & editing.

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Correspondence to Ganggang Zhang or Weihua Lai.

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Jing, X., Yu, S., Zhang, G. et al. Sensitive fluorescence ELISA for the detection of zearalenone based on self-assembly DNA nanocomposites and copper nanoclusters. Anal Bioanal Chem 416, 983–992 (2024). https://doi.org/10.1007/s00216-023-05088-6

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