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Rapid Homogeneous Detection of Biological Assays Using Magnetic Modulation Biosensing System

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Magnetic Nanoparticle-Assisted Multiplex RPA for Rapid Meat Authentication via Fluorescence Detection.

Cuiling Li¹, Zhen Wu¹, Daodong Pan¹

¹State Key Laboratory for Quality and Safety of Agro-Products, Zhejiang Key Laboratory of Intelligent Food Logistic and Processing, Zhejiang-Malaysia Joint Research Laboratory for Agricultural Product Processing and Nutrition and College of Food Science and Engineering, Ningbo University, Ningbo 315800, China.

Journal of Agricultural and Food Chemistry

|August 27, 2025

Summary

A new magnetic nanoparticle-assisted multiplex recombinase polymerase amplification (mRPA) assay rapidly detects horse and chicken in meat. This sensitive, field-deployable method ensures food authenticity and safety.

Keywords:

fluorescence detection high-throughput DNA isolation magnetic separation meat adulteration multiplex recombinase polymerase amplification

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Area of Science:

Food Science
Analytical Chemistry
Biotechnology

Background:

Meat adulteration with undeclared species is a significant global issue.
Accurate and rapid detection methods are crucial for food safety and regulatory compliance.

Purpose of the Study:

To develop a rapid, sensitive, and field-deployable assay for simultaneous detection of horse and chicken in meat.
To establish a high-throughput method for meat authentication and routine surveillance.

Main Methods:

Magnetic nanoparticle-assisted multiplex recombinase polymerase amplification (mRPA) with fluorescence detection.
Utilized streptavidin-coated Fe3O4@SiO2 nanoparticles and dual-labeled primers targeting ATP6-8 (horse) and CYTB (chicken) genes.
Integrated magnetic separation (MS) for amplicon isolation, reducing background noise and enhancing specificity.

Main Results:

The assay achieved a limit of detection of 0.1% (w/w) adulteration without cross-reactivity.
A 96-sample workflow, including DNA extraction, was completed in 31.5 minutes.
The mRPA-MS-FD assay demonstrated full agreement with national-standard RT-qPCR in analyzing 102 commercial samples, identifying 14 positives.

Conclusions:

The developed mRPA-MS-FD platform offers a sensitive, multiplexed, and high-throughput solution for meat authentication.
This assay is suitable for routine surveillance and point-of-need screening, addressing the pervasive problem of meat adulteration.