Rapid and Accurate Detection With CRISPR-Cas12b Based on Newly Identified -Specific and -Conserved Genomic Signatures.

is among the most prevalent foodborne zoonotic pathogens leading to diarrheal diseases. In this study, we developed a CRISPR-Cas12b-based system to rapidly and accurately detect contamination. Identification of -specific and -conserved genomic signatures is a fundamental step in development of the detection system. By comparing genome sequences with those of the closely related , followed by comprehensive online BLAST searches, a 20-bp -conserved (identical in 1024 out of 1037 analyzed genome sequences) and -specific (no identical sequence detected in non- strains) sequence was identified and the system was then assembled. In further experiments, strong green fluorescence was observed only when DNA was present in the system, highlighting the specificity of this system. The assay, with a sample-to-answer time of ∼40 min, positively detected chicken samples that were contaminated with a dose of approximately 10 CFU per gram of chicken, which was >10 times more sensitive than the traditional isolation method, suggesting that this method shows promise for onsite detection. This study provides an example of bioinformatics-guided CRISPR-Cas12b-based detection system development for rapid and accurate onsite pathogen detection.