Objective To develop a novel fluorescence polarization (FP) detection strategy and achieve highly sensitive Salmonella detection by incorporating recombinase polymerase amplification (RPA) technology.Methods The detection mechanism relies on homologous sequence recognition between target DNA and specific primers: When Salmonella targets exist, primer and target DNA binding triggers strand displacement and initiates DNA amplification. To enhance sensitivity, two key design features are implemented: 5'-end modification of primers with 6-carboxyfluorescein (6-FAM) as a fluorescent reporter, and selective digestion of unbound primers by exonuclease Ⅰ (Exo Ⅰ).Results During RPA amplification, the substantially increased molecular weight of amplicons restricts the rotational freedom of 6-FAM-labeled DNA compounds, resulting in significantly enhanced FP signals. After optimization, this method achieves a detection limit of 11 CFU/mL for Salmonella with excellent specificity.Conclusion This method provides an efficient Salmonella detection. Its modular design principle can also be readily adapted for rapid diagnosis of other foodborne pathogens.