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A Reciprocal-Amplifiable Fluorescence Sensing Platform via Replicated Hybridization Chain Reaction for Hosting Concatenated Multi-Ag Nanoclusters as Signal Reporter.

Mengdie LiJiayang HeXin ShangChunli YangYuqing ZhangSiyu ZuoRuo YuanWen-Ju Xu
Published in: Analytical chemistry (2022)
Exploring the replication of hybridization chain reaction HCR ( r HCR) for reciprocal amplification is intriguing in biosensing and bioanalysis. Herein, we develop a r HCR-based fluorescence platform that is manipulated by the combination of a specific DNA trigger ( T ) and a T -analogous amplicon ( T* ), thereby concatenating multi green-emissive Ag nanoclusters ( mg AgNCs) for amplifiable signal readout. Four well-designed hairpins (H1 recognizing T , H2, H3, and H4) with sequential complements are executed to operate r HCR. The termini of H1/H3 are merged to hybridize an inhibiting strand ( I ). The parent scaffold for mg AgNCs is separated into two splits (C 4 AC 4 T and C 3 GT 4 ) that are individually overhung in H2/H4. The presence of T activates the first HCR amplifier through cross-hybridization of four reactive hairpins for forming HCR duplexes. The next invasion of a complex ( T * ·I ) drives I to hybridize the tandem repeats in H1/H3, so that the displaced T* functions as T to catalyze the second amplifier r HCR for feeding back more hairpin assemblies with rapid reaction kinetics. In the shared r HCR polymers, the parent scaffolds (C 4 AC 4 TC 3 GT 4 ) in H2/H4 are collectively concatenated for the preferential clustering of mg AgNCs adducts, which cooperatively emit enormous T -responsive fluorescence signal. Because of the localization of T in HCR products, an alternative amplicon T* is introduced to drive r HCR progress via DNA strand displacement, generating more nucleating sites of emitters. Thus, the rational combination of nonenzymatic r HCR and label-free fluorescent concatemers would create a reciprocal signal amplification, achieving a simplified, rapid, and highly sensitive assay down to femtomolar concentrations.
Keyphrases
  • label free
  • single molecule
  • nucleic acid
  • quantum dots
  • high throughput
  • energy transfer
  • cell free
  • drug delivery
  • sensitive detection
  • mass spectrometry
  • highly efficient
  • liquid chromatography
  • simultaneous determination