The adulteration of food products with porcine components is among the most widespread concerns in the global food industry. Apart from health and religious reasons, being able to detect porcine adulteration is crucial to warrant fair trading and legal compliance. Current detection methods are often constrained by their low sensitivity and expensive equipment. In this work, a recombinase polymerase amplification (RPA) assay for the rapid and sensitive detection of porcine is constructed, with propidium iodide (PI) dye as the reporter molecule. Under the optimized conditions, the RPA-PI assay achieved a wide linear range of 0.005–100 ng/μL and a detection limit of 0.005 ng/μL for porcine DNA with no cross-reactivity. Furthermore, the application of the assay to actual food samples demonstrated a 100 % consistency with conventional PCR results. This developed assay not only provides a novel porcine adulteration detection platform with simpler and faster protocols, but it also holds the potential to be developed further for point-of-care authenticity testing.
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