Concatenated Catalytic Hairpin Assembly/Hyperbranched Hybridization Chain Reaction Based Enzyme-Free Signal Amplification for the Sensitive Photoelectrochemical Detection of Human Telomerase RNA

Human telomerase RNA (hTR), an important biomarker for cancer diagnosis, is the template for the synthesis of telomeric DNA repeats and is found to be 7-fold overexpressed in tumor cells. Herein, we present a photoelectrochemical (PEC) biosensor for hTR detection coupled with a novel amplification strategy based on cascades of catalytic hairpin assembly (CHA) and hyperbranched hybridization chain reaction (HB-HCR). At the electrode surface, thiolated hairpin 1 probes were immobilized on deposited CdS nanoparticles via a Cd–S bond. In the presence of target hTR, a CHA reaction was triggered and the exposing of trigger1 could further initiate an HB-HCR reaction to form abundant hemin/G-quadruplex DNAzymes containing dendritic DNA structure. The DNAzymes’ catalytic precipitation of 4-chloro-1-naphthol (4-CN) by H2O2 subsequently took place on the surface of the PEC electrode and efficiently suppressed the photocurrent output. Therefore, the change of photocurrent response had a positive linear relationship with logarithmic value of hTR concentration varying from 200 fM to 20.0 nM with a limit of detection (LOD) of 17.0 fM. The LOD for CHA/HB-HCR was about 8.8-fold lower than that of CHA/linear-branched HCR (CHA/LB-HCR) and 547-fold lower than that of CHA. By coupling the feature of high signal amplification capacity for DNA nanotechnology, a prominently stable, reproducible, and selective PEC biosensor was successfully constructed and applied in hTR detection.