Trace detection of S. aureus cells in food samples via RCA-assisted SERS signal amplification with core-shell...

Staphylococcus aureus (S. aureus) has been identified as a indicator of food contamination. In this study, a sensitive and accurate biosensor strategy for S. aureus through rolling circle amplification-assisted surface-enhanced Raman scattering (RCA-assisted-SERS), has been established. The work relies on the interaction between the aptamer and its partial complementary DNA strands fabricated on the surface of gold and silver-assisted magnetic microspheres and the subsequent detachment to trigger the activation of the RCA process. In RCA, template DNA, T4 DNA ligase, and Phi29 DNA polymerase were assembled to form long single-stranded DNA containing repetitive sequences. The gold core encapsulated with a layer of 4-nitrothiophenol and further covered with a silica shell was employed as the SERS nanoprobe (Au@NTP@SiO₂). Subsequently, the output and amplification of SERS signal were performed by hybridizing ssDNA functionalized Au@NTP@SiO₂ to realize the quantitative detection of S. aureus. Under the optimal conditions, S. aureus sensing was monitored (36.0-3.6 × 10⁸ cfu/mL) with a limit of detection of 2.0 cfu/mL. This strategy was further validated for S. aureus recognition in spiked real samples with favorable recoveries (94.0-103.4 %) at p > 0.05. The suggested RCA-assisted SERS approach exhibits potential for multiple foodborne pathogens in both food safety and biomedical investigations.