Foodborne
pathogens can cause illnesses. Existing tools for detecting
foodborne pathogens are typically time-consuming or require complex
protocols. Here, we report an assay to directly analyze pathogenic
genes based on CRISPR-Cas12. This new test, termed proximal DNA probe-based
CRISPR-Cas12 (PPCas12), facilitates the detection of foodborne pathogens
without amplification steps. The elimination of the nucleic acid amplification
process dramatically reduced the processing time, complexity, and
costs in the analysis of foodborne pathogens. The substitution of
the frequently used dually labeled DNA reporter with a proximal DNA
probe in the PPCas12 assay led to a 4-fold sensitivity enhancement.
PPCas12 offered a limit of detection of 619 colony-forming units in
the detection of Salmonella enterica (S. enterica) without the nucleic
acid amplification process. The specific recognition of genes via
PPCas12 allowed distinguishing S. enterica from other foodborne pathogens. The PPCas12 assay was applied in
the screening of S. enterica contamination
on fresh eggs with high precision. Hence, the new PPCas12 assay will
be a valuable tool for on-site monitoring of foodborne pathogens.
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