Recombinase polymerase amplification combined with Pyrococcus furiosus Argonaute for fast Salmonella spp. testing in food safety

Int J Food Microbiol. 2024 Jun 2:417:110697. doi: 10.1016/j.ijfoodmicro.2024.110697. Epub 2024 Apr 16.

Abstract

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 (PfAgo)-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.

Keywords: Foodborne pathogens; PfAgo; Recombinase polymerase amplification; Salmonella spp. testing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Argonaute Proteins / genetics
  • Argonaute Proteins / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Food Contamination / analysis
  • Food Microbiology*
  • Food Safety
  • Nucleic Acid Amplification Techniques* / methods
  • Pyrococcus furiosus* / genetics
  • Recombinases / genetics
  • Recombinases / metabolism
  • Salmonella* / genetics
  • Salmonella* / isolation & purification
  • Sensitivity and Specificity

Substances

  • Recombinases
  • Bacterial Proteins
  • Argonaute Proteins
  • invA protein, Bacteria