Portable biosensor for quantitative detection of meat adulteration based on heparin sodium-mediated one-tube RPA/SCas12a amplification strategy

Meat adulteration poses significant social concerns, including the infringement of consumer rights, potential risks of food allergies, and conflicts with religious dietary practices. Therefore, a pressing need exists for the creation of swift and highly sensitive techniques to verify the authenticity of meat products. A portable and quantitative detection assay for pork identification in food was established by integrating a personal glucose meter (PGM) with a heparin-mediated one-pot RPA/Cas12a (called hRPA/SCas12a-PGM). In the hRPA/SCas12a-PGM assay, the target DNA sequence initiates the heparin-enhanced RPA/Cas12a reaction in a single tube, subsequently activating Cas12a’s DNase function. Once activated, Cas12a cleaves sucrase-labeled DNA probes immobilized on the electrode, which are labeled with sucrase. Sucrase-labeled DNA fragments are released into the solution through this cleavage process. Sucrase then catalyzes the conversion of sucrose into glucose, producing a quantifiable signal that is detected by the PGM. The integrated system exhibited a broad dynamic range (10 pg/μL to 100 ng/μL), a low detection limit (10 pg/μL), and strong specificity against non-target samples. The hRPA/SCas12a-PGM assay provides a powerful, field-deployable solution for meat authenticity testing, offering significant potential for application in food safety surveillance and regulatory enforcement. Graphical