A one-pot and one-step RPA-CRISPR/Cas12b platform applied for streamlined detection of Salmonella spp.

Salmonella spp. remains a primary culprit behind foodborne illnesses, posing significant risks to human health and the economy. Existing detection methods for Salmonella are often hindered by several drawbacks. Here, a streamlined and precise detection approach was introduced for Salmonella , leveraging the integration of RPA technology with the CRISPR/Cas12b system. This combined RPA-CRISPR/Cas12b method operates in a single PCR tube, simplifying the workflow to a one-step process. Our findings indicate that this assay has a limit of detection (LOD) of 60 copies per test and can be completed within just one hour. Importantly, the assay demonstrated 100% specificity for Salmonella , without any cross-reactivity with other bacterial strains. In practical tests, positive amplification was achieved even in the case that Salmonella contamination level reached 19.5 CFU/ml. Furthermore, an equal number of positive poultry samples were detected by using traditional culture method and the present RPA-CRISPR/Cas12b method, including chicken (52/483, 28.42%), duck (19/40, 47.50%), goose (14/31, 45.16%), and pigeon (10/52, 19.23%), respectively. Overall, this research introduces a novel detection system for Salmonella and underscores the promising potential of this method in curbing foodborne diseases.