Rapid and accurate detection of Salmonella enterica serovar Enteritidis using a one-step LAMP-CRISPR/Cas12b method

Salmonella enterica serovar Enteritidis (S. Enteritidis) poses a significant threat as a prevalent foodborne pathogen globally. Timely diagnostic methods are essential for effective management of S. Enteritidis infections. Herein, we establish a one-step LAMP-CRISPR/Cas12b method for a rapid, accurate, and convenient detection of S. Enteritidis. The entire reaction requires only 45 min without the need for specialized equipment. The platform exhibited 100% specificity with no cross-reactivity observed among 19 other Salmonella serovars and 9 non- Salmonella bacterial pathogens. Our method achieved a limit of detection of 30 copies per reaction. Compared with the traditional culture methods in practical analysis. S. Enteritidis positivity rates identified by one-step LAMP-CRISPR/Cas12b platform were 2.88% (6/208) including 3.14% (4/127) in chicken, 4.44% (2/45) in duck, and 0.0% (0/36) in pork, and a 100% concordance was observed between the traditional culture method and one-step LAMP-CRISPR/Cas12b platform. In summary, our findings underscore the simplicity and accuracy of the one-step LAMP-CRISPR/Cas12b platform for S. Enteritidis detection, with potential applications in resource-limited settings.