Sensitive Detection of a Single-Nucleotide Polymorphism in Foodborne Pathogens Using CRISPR/Cas12a-Signaling ARMS-PCR

Abstract

<i>Salmonella</i> infection, particularly that caused by drug-resistant strain, has become a worldwide public health issue. Herein, we presented a CRISPR/Cas12a-signaling ARMS-PCR assay, termed cARMS, capable of sensitively detecting drug-resistant <i>Salmonella enterica</i> (<i>S. enterica</i>) involving single-nucleotide polymorphism (SNP). Owing to the dual-recognition processes, i.e., allele-specific primed polymerization and CRISPR/Cas12 binding, the cARMS assay yielded a high sensitivity for detecting SNP down to ∼0.5%. We used the cARMS assay to investigate the adaptation of SNP-involved drug-resistant <i>S. enterica</i> to salt stress. It was found that the mutants exhibited stronger adaptation to salt stress, indicating the potential risk of using high salt content as a sterilization strategy. The results verified the feasibility of the cARMS assay in controlling SNP-involved bacteria-associated biosafety.