Competitive Ratiometric Aptasensing with Core-Internal Standard-Shell Structure Based on Surface-Enhanced Raman Scattering.

Reproducibility and stability are important indicators for the evaluation of quantitative sensing methods based on surface-enhanced Raman scattering (SERS) technology. Developing a SERS substrate with self-calibration capabilities is vital for effectively quantifying targets. In this work, a competitive ratiometric SERS aptasensor was developed. 4-Aminothiophenol as an internal standard (IS) was embedded in the substrate followed by gradually loading with the aptamer and methylene blue functionalizing of the complementary sequences of the aptamer (MB-cDNA). Recognition and binding of the target to the aptamer resulted in the shedding of MB-cDNA after magnetic separation reducing the SERS signal of MB, allowing for the ratiometric determination of the target based on the constant intensity from the IS. For the selective detection of okadaic acid (OA), a good negative correlation was achieved between the SERS ratiometric intensity and OA concentration in the range of 0.5-100 ng/mL. The magnetic separation strategy effectively simplifies the production steps of the aptasensor, and the ratiometric strategy effectively improved the reproducibility and stability of the OA sensing. This ratiometric aptasensor has been successfully employed to detect OA in food and environmental samples and is expected to be extended to detect other targets.