Search

Foodborne illness caused by Salmonella spp. is one of the most prevalent public health problems globally, which have brought immeasurable economic burden and social impact to countries around the world. Neither current nucleic acid amplification detection method nor standard culture method (2–3 days) are suitable for field detection in areas with a heavy burden of Salmonella spp. Here, we developed a highly sensitive and accurate assay for Salmonella spp. detection in less than 40 min. Specifically, the invA gene of Salmonella spp. was amplified by recombinase polymerase amplification (RPA), followed by Pyrococcus furiosus Argonaute (Pf Ago)-based target sequence cleavage, which could be observed by a fluorescence reader or the naked eye. The assay offered the lowest detectable concentration of 1.05 × 101 colony forming units/mL (CFU/mL). This assay had strong specificity and high sensitivity for the detection of Salmonella spp. in field samples, which indicated the feasibility of this assay. • The method can be widely used in on-site food sample detection or standard laboratory detection. • The operation process is simple and fast (∼40 min). • High detection sensitivity • Ultraviolet light can read signals. [ABSTRACT FROM AUTHOR]

At present, the main challenge for Staphylococcus aureus (S. aureus) in food safety is still a lack of rapid, convenient detection methods. This study developed a combination of recombinase polymerase amplification (RPA) and the CRISPR-Cas12a technology (known as the RPA-CRISPR-Cas12a assay) for the detection of S. aureus in food. This novel assay could detect samples with a fluorescence reader, lateral flow dipstick (LFD) or the naked eye under a UV transilluminator. After optimization, the RPA-CRISPR-Cas12a assay could specifically and stably detect S. aureus as low as 1–10 copies/μL within 40 min at 37 °C. This assay had high sensitivity to the detection of S. aureus in both laboratory standard samples and field samples, which indicated the feasibility of this assay. To sum up, this accurate, portable detection method for S. aureus has great potential applications in food safety, especially in the field, without requiring sophisticated equipment or high level of technical expertise. [ABSTRACT FROM AUTHOR]