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Programmable Hairpin Argonaute-Nanopore Detection System for Multiplex Mycotoxin Analysis
Zhuoqun Su1, Ziye Pei1, Jianing Chen1
1School of Food Science and Engineering, Shaanxi University of Science and Technology, Xi'an 710021, China.
Analytical Chemistry
|October 16, 2025
Summary
A new sensing platform, programmable hairpin argonaute-nanopore detection (PHAND), enables simultaneous detection of multiple food contaminants. This method uses Thermus thermophilus Argonaute (TtAgo) and DNA probes for sensitive and specific analysis.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Food Science
Background:
- Conventional methods struggle with simultaneous detection of multiple food contaminants.
- Lack of accurate multiplexed signal conversion hinders current sensing technologies.
Purpose of the Study:
- To develop a versatile sensing platform for multiplexed detection of small-molecule hazardous substances.
- To enable portable and rapid food safety screening.
Main Methods:
- Integration of Thermus thermophilus Argonaute (TtAgo)-mediated cleavage with DNA hairpin probes.
- Utilizing guide DNA release to activate TtAgo for specific cleavage of hairpin probes.
- Generating distinct nanopore reporters that produce orthogonal current blockade signals via alpha-hemolysin nanopores.
Main Results:
- Successful simultaneous detection of three mycotoxins: aflatoxin B1, ochratoxin A, and zearalenone.
- Demonstrated high sensitivity and specificity in detecting target analytes.
- Validated the method in spiked real food samples with satisfactory recovery rates.
Conclusions:
- The PHAND system offers a novel approach for portable and multiplexed detection of hazardous substances in food.
- This technology holds significant potential for rapid and reliable food safety screening.
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