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Updated: May 29, 2026
Species Determination and Quantitation in Mixtures Using MRM Mass Spectrometry of Peptides Applied to Meat Authentication
Published on: September 20, 2016
Universal primer-based RPA combined with parallel CRISPR/Cas12a decoding for rapid multi-species meat authentication
Shuang Han1, Xinyi Lin1, Yuting Lei1
1Faculty of Forensic Medicine, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, 510080 Guangzhou, PR China; Guangdong Province Translational Forensic Medicine Engineering Technology Research Center, Sun Yat-sen University, Guangzhou 510080, PR China.
A new rapid assay uses universal primers and CRISPR/Cas12a technology for quick meat species identification. This cost-effective method accurately detects 11 meat types and identifies adulteration in food products.
Area of Science:
- Food Science
- Molecular Biology
- Biotechnology
Background:
- Accurate meat authentication is crucial for ensuring food safety and preventing economic fraud.
- Existing methods for meat species identification can be time-consuming, expensive, or require specialized equipment.
- There is a need for rapid, specific, and cost-effective tools for detecting multiple meat species and adulteration.
Purpose of the Study:
- To develop and validate a novel two-step assay for the rapid identification of 11 common meat species.
- To assess the assay's specificity, sensitivity, and ability to detect adulteration in meat mixtures.
- To provide a cost-effective and efficient platform for meat authentication.
Main Methods:
- Utilized a two-step approach combining universal primer-based Recombinase Polymerase Amplification (RPA) with species-specific CRISPR/Cas12a detection.
- Employed a single universal primer pair for broad DNA amplification, followed by parallel single-target CRISPR reactions using specific crRNAs.
- Integrated dual fluorescence and lateral flow strip detection formats for versatile readout.
Main Results:
- The assay successfully identified 11 common meat species with high specificity and no cross-reactivity.
- Detection limits ranged from 10^0 to 10^4 copies/μL, demonstrating high sensitivity.
- The fluorescence assay detected adulteration down to 0.05% (w/w), while the lateral flow format detected down to 0.05-5% (w/w), depending on the species.
- The assay was validated using commercially processed meat products.
Conclusions:
- This laboratory-validated strategy offers a rapid (approx. 40 min), cost-effective (∼$4 per test), and specific method for identifying multiple meat species.
- The simplified primer design and parallel detection capabilities make it a promising platform for qualitative meat screening.
- Further validation is needed to confirm its robustness for field applications.
