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

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.

 

Comments:

The process you've described is quite fascinating! It seems like a sophisticated approach to enhance the sensitivity of detecting Zearalenone (ZEN), a mycotoxin produced by Fusarium species, which poses potential risks to human health if present in food.

Let me break it down a bit:

1. **Challenges with Traditional ELISA:** The conventional ELISA method faces limitations in sensitivity when detecting ZEN.

2. **Enzyme Nanocomposites (ASBD):** To overcome these limitations, enzyme nanocomposites (ALP-SA-Bio-ssDNA, ASBD) were created using streptavidin-labeled alkaline phosphatase (SA-ALP) and dual-biotinylated single-stranded DNA (B2-ssDNA). These nanocomposites allowed for a higher loading amount of ALP, which increased the conversion of ascorbic acid 2-phosphate to generate ascorbic acid (AA).

3. **Formation of Copper Nanoclusters (CuNCs):** Ascorbic acid (AA) was then used to reduce Cu2+ to form copper nanoclusters (CuNCs) by interacting with poly T. These CuNCs emit a strong fluorescence signal.

4. **Fluorescence ELISA Development:** By leveraging the high enzyme load of the nanocomposites and the strong fluorescence signal of CuNCs, a fluorescence-based ELISA method was successfully established for detecting ZEN.

5. **Enhanced Sensitivity:** This new method demonstrated a lower limit of detection (0.26 ng mL-1) compared to the traditional ELISA (1.55 ng mL-1), indicating significantly improved sensitivity.

6. **Accuracy and Selectivity:** The method's accuracy was verified through recovery rates ranging from 92.00% to 108.38%, with low variation coefficients (< 9.50%) when detecting ZEN in corn and wheat samples. Moreover, it exhibited no cross-reaction with four other mycotoxins, ensuring its specificity for ZEN detection.

7. **Applications:** The enhanced sensitivity and specificity make this method highly promising for trace detection in food safety protocols.

This innovative approach could significantly impact food safety measures by enabling more accurate and sensitive detection of ZEN, thereby reducing potential health risks associated with its presence in food products.

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