Proximity hybridization-triggered cascade amplification for label-free SERS detection of Alzheimer's amyloid-β oligomers

Most of the existing SERS systems failed to achieve satisfactory results in early diagnosis of Alzheimer's disease owing to a lack of effective signal transduction. Herein, we developed a dual signal amplification strategy for SERS detection of amyloid-beta oligomers based on proximity hybridization-triggered catalyzed hairpin assembly (CHA) and hybridization chain reaction (HCR). In the presence of the target protein and two DNA-labeled antibodies, a proximate complex formed in a homogeneous solution. Each of the A beta O-DNA complexes served as a catalyst to trigger and accelerate numerous hybridization processes between MB1 and MB2. Subsequently, the single-strand fragment on the electrode surface initiated HCR, resulting in the hybridization reaction to form double-strand DNA concatemers on the substrate surface. The surface became negatively charged and allowed the absorption of silver ions on the DNA skeleton. After chemical reduction by hydroquinone, the formed silver nanoparticles could be further grown with a silver enhancement step to amplify the detectable SERS signal by absorbing rhodamine 6G as a SERS reporter on the silver nanoparticle surface. This biosensing platform had potential applications in molecular diagnostics of AD serum samples.