Abstract
Meat adulteration poses serious economic, regulatory, and ethical challenges worldwide, creating an urgent need for rapid, on-site, and multiplex authentication methods. Although polymerase chain reaction (PCR) is the gold standard for nucleic acid analysis, its reliance on laboratory infrastructure and skilled operators limits field deployment, while current isothermal amplification platforms still lack sufficient automation and integration. Here, we present Magtect, a fully automated magnetofluidic system for rapid, multiplex identification of meat adulteration in sheep products. The system integrates magnetic bead-based nucleic acid extraction, ultrasound-assisted washing, magnetic array-guided bead distribution, and parallel multiplex recombinase polymerase amplification (RPA) detection within a single chip. By combining silicone oil with a thermally controllable wax isolation layer, Magtect enables physical spatial separation of multi-step reagents and on-demand mixing during heating, effectively preventing premature reagent contact and cross-interference. Notably, the wax barrier isolates the elution buffer from the RPA master mix, eliminating bead-induced amplification inhibition commonly observed in conventional magnetofluidic designs. Using this platform, adulteration of sheep meat with duck, chicken, or pork components can be fully automatically identified within 30 min. The system achieves a detection limit of 0.1 copies/μL and reliably detects adulteration levels as low as 1%. These results demonstrate that Magtect provides a robust, sensitive, and field-deployable solution for on-site meat authenticity verification, representing a significant step forward in automated food integrity monitoring.