Exploring the replication of hybridization
chain reaction HCR (rHCR) for reciprocal amplification
is intriguing in biosensing
and bioanalysis. Herein, we develop a rHCR-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
(mgAgNCs) for amplifiable signal readout. Four well-designed
hairpins (H1 recognizing
T
, H2, H3,
and H4) with sequential complements are executed to operate rHCR. The termini of H1/H3 are merged to hybridize an inhibiting
strand (I). The parent scaffold for mgAgNCs is separated into two splits (C4AC4T
and C3GT4) 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 rHCR for feeding
back more hairpin assemblies with rapid reaction kinetics. In the
shared rHCR polymers, the parent scaffolds (C4AC4TC3GT4) in H2/H4 are collectively
concatenated for the preferential clustering of mgAgNCs 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 rHCR progress via DNA strand displacement, generating more
nucleating sites of emitters. Thus, the rational combination of nonenzymatic rHCR and label-free fluorescent concatemers would create
a reciprocal signal amplification, achieving a simplified, rapid,
and highly sensitive assay down to femtomolar concentrations.