Detection and absolute quantification biosensing tools for food authentication: CRISPR/Cas, digital CRISPR and beyond

Food authenticity is essential to protect against fraudulent activities and ensure food safety. However, traditional authentication methods have limited capabilities due to lengthy testing period, bulky equipment, and low sensitivity and specificity. To address this, CRISPR/Cas and digital CRISPR (dCRISPR) have emerged as potential tools for quick and reliable food authentication. This review focuses on the CRISPR/Cas system and absolute quantification technologies, like sample partitioning-based dCRISPR, for food authentication. It provides a detailed overview of various Cas proteins for detection, diverse amplification methods to enhance signal, and different digital sample partitioning techniques facilitating absolute quantification. Notably, it explores the latest advancements of CRISPR/Cas and digital assays for food authentication applications. Furthermore, it discusses the potential opportunities and challenges that CRISPR/Cas and dCRISPR present in food authentication. CRISPR/Cas and dCRISPR are ground breaking technologies for food authentication. When combined with digital sample partitioning, dCRISPR become even more sensitive and accurate than normal CRISPR/Cas-based detection, while also providing absolute quantification. These techniques can detect and quantify food adulterants, identify genetically modified food, distinguish closely-related species, and authenticate clean labels. With improved performance, CRISPR/Cas and dCRISPR have the potential to revolutionize the food supply chain.