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In this work, single-stranded DNA aptamers that are highly specific to enterotoxigenic Salmonella paratyphi A were obtained from an enriched oligonucleotide pool using Systematic Evolution of Ligands by Exponential Enrichment (SELEX) to target the flagellin protein. The selected aptamers were confirmed to have high sensitivity and specificity to the flagellin. In addition, a probe (P0) containing the DNAzyme and fluorescein isothiocyanate-labeled aptamer3 sequences was employed as a dual probe for observing fluorescence and absorbance changes. The flagellin demonstrated low detection limits of 5 ng/mL by fluorescence and 20 ng/mL by spectrophotometry. Moreover, milk samples spiked with Salmonella paratyphi A were also detected, with the low detection limits increasing to 10 5 CFU/mL by fluorescence and 10 6 CFU/mL by spectrophotometry. The combination of fluorescence and spectrophotometry offers a specific, rapid, and sensitive way for detecting Salmonella paratyphi A and has potential for detecting other pathogens in food.

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