Duplex nanoplate-based digital PCR assay for simultaneous detection of pork and chicken in processed meat products

In this study, we developed a novel molecular detection assay for the simultaneous identification of pork and chicken in processed foods using nanoplate-based digital PCR. This assay, which targets the mitochondrial D-loop region and the cytochrome b gene, demonstrated high specificity against 11 animal species. Sensitivity was assessed using tenfold serial dilutions of DNA, and the results showed comparable sensitivity to real-time PCR. Notably, the limit of detection (LOD) of this assay was 0.1 % (w/w) for both pork and chicken in beef, which was more sensitive than the 1 % (w/w) LOD observed with real-time PCR. We further evaluated its applicability using 27 commercial meat products, confirming that the results were consistent with the species information provided on the labels. Therefore, this novel duplex nanoplate-based digital PCR assay offers a practical solution for the accurate detection of pork and chicken in various meat-based matrices, highlighting its potential for applications in the food industry.