The authors describe an aptamer-based assay for 17β-estradiol. It relies on the combined use of surface enhanced Raman scattering (SERS) and hybridization chain reaction (HCR). The aptamer against 17β-estradiol is applied as the recognition probes, and this results in excellent specificity. Specific recognition of target 17β-estradiol induce the freedom of DNA 2, which will open the stem-loop structure of probe 1 on the Au@Ag and form the partial dsDNA structure. With the nicking enzyme, the partial dsDNA will be hydrolyzed and the reside ssDNA on Au@Ag will form a small stem-loop structure. With the help of the other probe 2 modified Au@Ag and pre-immobilized probe 3 on the well of the microplate, an enzyme-free HCR can occur and tremendous Au@Ag can be assembled along the formed dsDNA in HCR, which can act as the excellent substrate for Raman measurement and greatly amplify the Raman signal of R6G on the Au@Ag. Afterwards, the key factor, ratio between probe 2-Au@Ag (P2) and probe1-Au@Ag (P1), affects the detection sensitivity is systematically optimized for the best sensing performance. The SERS signal of R6G, best measured at 1651 cm-1, increases linearly in the wide range from 1 pM to 10 nM. The detection limit can be as low as 0.1 pM. Graphical abstract Schematic presentation of an aptamer-based surface enhanced Raman scattering method for accurate detection of 17β-estradiol, which is integrated with hybridization chain reaction for signal amplification and sensitivity improvement.
Keywords: Aptamer; Environment monitoring; Estrogen; Food safety; Gold nanoparticle; Hybridization chain reaction; SERS; Signal amplification.