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Monoclonal antibody-quantum dots CdTe conjugate-based fluoroimmunoassay for the determination of aflatoxin B1 in
Zhaowei Zhang1, Yuanyuan Li, Peiwu Li
1Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences, Wuhan 430062, PR China; Key Laboratory of Detection for Mycotoxins, Ministry of Agriculture, Wuhan 430062, PR China; Laboratory of Risk Assessment for Oilseeds Products (Wuhan), Ministry of Agriculture, Wuhan 430062, PR China.
Insights
This study presents a new fluoroimmunoassay using quantum dots for detecting aflatoxin B1 (AFB1) in peanuts. The method offers rapid, sensitive, and accurate AFB1 quantification, with potential for multiplexed mycotoxin analysis.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Materials Science
Background:
- Aflatoxin B1 (AFB1) is a toxic secondary metabolite produced by Aspergillus species, posing significant health risks.
- Accurate and sensitive detection methods for AFB1 are crucial for food safety and regulatory compliance.
- Existing detection methods may lack the sensitivity, speed, or multiplexing capability required for comprehensive analysis.
Purpose of the Study:
- To develop a novel fluoroimmunoassay for the sensitive detection of aflatoxin B1 (AFB1).
- To utilize quantum dots (QDs) as fluorescent labels for enhanced assay performance.
- To establish a direct competitive fluorescence-linked immunosorbent assay (cFLISA) for AFB1 quantification in food matrices.
Main Methods:
- Conjugation of Cadmium Telluride (CdTe) quantum dots with monoclonal antibodies specific to AFB1.
- Development of a direct competitive fluorescence-linked immunosorbent assay (cFLISA) protocol.
- Validation of the cFLISA method using spiked peanut samples to assess performance parameters.
Main Results:
- The developed cFLISA demonstrated a 50% inhibition concentration (IC50) of 0.149 ng/mL in a peanut matrix.
- The limit of detection (LOD) was determined to be 0.016 ng/mL.
- High recovery rates (85-117%) and low coefficients of variation (<10%) were achieved, indicating method accuracy and precision.
Conclusions:
- The novel QD-based cFLISA offers a rapid, sensitive, and reliable method for AFB1 detection in peanuts.
- This assay provides a valuable alternative to existing methods for AFB1 quantification.
- The platform shows significant potential for simultaneous determination of multiple mycotoxins, enhancing analytical throughput.
Abstract:
A fluoroimmunoassay towards aflatoxin B1 (AFB1) was presented using quantum dots as the fluorescent label. The CdTe QDs were successfully linked to the monoclonal antibody against AFB1. Based on the conjugated complexes, a novel direct competitive fluorescence-linked immunosorbent assay (cFLISA) was developed for AFB1 detection. The 50% inhibition value (IC50) of the cFLISA was 0.149ng/mL in peanuts matrix. The method performance included the limit of detection (LOD) of 0.016ng/mL and considerable recoveries of 85-117% at three fortification levels (0.075, 0.15, and 0.3ng/g) from spiked AFB1 blank peanuts samples, along with coefficients of variation (CVs) below 10%. The cFLISA provided an alternative of rapid and sensitive detection for AFB1 and, moreover provided great potential for multiplexed mycotoxins determination simultaneously.
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