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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.

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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.