The detection of Salmonella in food based on PCR combined with Pyrococcus furiosus Argonaute

Salmonella is a common zoonotic foodborne bacterial pathogen. In poultry farms, Salmonella can cause pullorum disease, which characterizes decreased appetite and diarrhea in chicks, accompanied by a high mortality rate in the first week of life itself. In pig farms, Salmonella mainly infects piglets and causes porcine paratyphoid, characterizing septicemia and necrotic enteritis and sometimes leading to severe symptoms such as encephalitis. This bacterium poses significant challenges to global public health and food safety. PfAgo (Pyrococcus furiosus Argonaute) is an Argonaute enzyme from the Pyrococcus furiosus, which is utilized in highly sensitive and accurate nucleic acid detection and analysis methods based on the polymerase chain reaction (PCR) combined with PfAgo. In this study, targeting the hilA gene, and can detect as low as 3 × 100 copies and 2.7 × 10°CFU (cells) with no cross-reactivity with other pathogens, it enhances the sensitivity of PCR. The PCR-PfAgo method was successfully applied to detect in clinical food samples including chicken and milk, and the results were consistent with those of traditional bacterial culture. The findings suggest that the PCR-PfAgo method has the potential to replace traditional detection methods and has broad application prospects in the detection of Salmonella pathogen.