LAMP assay coupled CRISPR/LbCas12a system in a single-tube method for visual detection of meat adulteration

Meat adulteration frequently infringes upon consumers' rights and interests, disrupts the market, and violates religious beliefs. However, implementing on-site detection of meat samples remains technically challenging for widespread applications. Here, we developed a point-of-care testing (POCT) platform for fast, highly sensitive, and on-site detection of meat adulteration, based on loop-mediated isothermal amplification (LAMP) coupled the clustered regularly interspaced short palindromic repeats (CRISPR) system in a single-tube. This platform integrates the efficient exponential amplification capacity of LAMP and the high specificity recognition capability of CRISPR, reducing the false positive results and minimizing LAMP aerosol contamination risk. Pork was selected as the target for adulteration detection, qualitative detection by the naked eye can be achieved within 1 h from sample collection to results with a sensitivity as high as 0.05 % to detect binary mixtures of pork in beef and the limit of DNA detection was at 50 pg/mu L which 10 times higher than the individual LAMP assay. This work is expected to offer a powerful universal POCT platform for addressing a range of meat adulteration issues, suitable for enhancing food safety measures.