Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array

Abnormal expression of miRNAs is associated with the occurrence and progression of cancer and other diseases, making miRNAs essential biomarkers for disease diagnosis and prognosis. However, the intrinsic properties of miRNAs, such as short length, low abundance, and high sequence homology, represen...

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Main Authors: Bin Wang, Huiqiang Ma, Mingxing Zhou, Xian Huang, Ying Gan, Hong Yang
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Journal of Functional Biomaterials
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Online Access:https://www.mdpi.com/2079-4983/16/3/98
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author Bin Wang
Huiqiang Ma
Mingxing Zhou
Xian Huang
Ying Gan
Hong Yang
author_facet Bin Wang
Huiqiang Ma
Mingxing Zhou
Xian Huang
Ying Gan
Hong Yang
author_sort Bin Wang
collection DOAJ
description Abnormal expression of miRNAs is associated with the occurrence and progression of cancer and other diseases, making miRNAs essential biomarkers for disease diagnosis and prognosis. However, the intrinsic properties of miRNAs, such as short length, low abundance, and high sequence homology, represent great challenges for fast and accurate miRNA detection in clinics. Herein, we developed a novel hybridization chain reaction (HCR)-based electrochemical miRNAs chip (e-miRchip), featured with gold nanostructured electrodes (GNEs) and silver nanoparticle reporters (AgNRs), for sensitive and multiplexed miRNA detection. AgNRs were synthesized and applied on the e-miRchip to generate strong redox signals in the presence of miRNA. The stem–loop capture probe was covalently immobilized on the GNEs, and was opened upon miRNA hybridization to consequently trigger the HCR for signal amplification. The multiple long-repeated DNA helix generated by HCR provides the binding sites for the AgNRs, contributing to the amplification of the electrochemical signals of miRNA hybridization. To optimize the detection sensitivity, GNEs with three distinct structures were electroplated, in which flower-like GNEs were found to be the best electrode morphology for miRNAs analysis. Under optimal conditions, the HCR-based e-miRchip showed an excellent detection performance with an LOD of 0.9 fM and a linear detection range from 1 fM to 10 pM. Moreover, this HCR-based e-miRchip platform was able to effectively distinguish miRNAs from the one- or two-base mismatches. This HCR-based e-miRchip holds great potential as a highly efficient and promising miRNA detection platform for the diagnosis and prognosis of cancer and other diseases in the future.
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spelling doaj-art-e71c166694a8480eaeeb3ca86e1f80072025-08-20T02:42:31ZengMDPI AGJournal of Functional Biomaterials2079-49832025-03-011639810.3390/jfb16030098Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode ArrayBin Wang0Huiqiang Ma1Mingxing Zhou2Xian Huang3Ying Gan4Hong Yang5Intensive Care Unit, Department of Emergency Medicine, The Second Hospital of Tianjin Medical University, Tianjin 300211, ChinaDepartment of Pharmacology and Tianjin Key Laboratory of Inflammation Biology, The Province and Ministry Co-Sponsored Collaborative Innovation Center for Medical Epigenetics, School of Basic Medical Sciences, Tianjin Medical University, No. 22 Qixiangtai Road, Heping District, Tianjin 300070, ChinaDepartment of Biomedical Engineering, Tianjin University, 92 Weijin Road, Tianjin 300072, ChinaDepartment of Biomedical Engineering, Tianjin University, 92 Weijin Road, Tianjin 300072, ChinaDepartment of Pharmacology and Tianjin Key Laboratory of Inflammation Biology, The Province and Ministry Co-Sponsored Collaborative Innovation Center for Medical Epigenetics, School of Basic Medical Sciences, Tianjin Medical University, No. 22 Qixiangtai Road, Heping District, Tianjin 300070, ChinaDepartment of Pharmacology and Tianjin Key Laboratory of Inflammation Biology, The Province and Ministry Co-Sponsored Collaborative Innovation Center for Medical Epigenetics, School of Basic Medical Sciences, Tianjin Medical University, No. 22 Qixiangtai Road, Heping District, Tianjin 300070, ChinaAbnormal expression of miRNAs is associated with the occurrence and progression of cancer and other diseases, making miRNAs essential biomarkers for disease diagnosis and prognosis. However, the intrinsic properties of miRNAs, such as short length, low abundance, and high sequence homology, represent great challenges for fast and accurate miRNA detection in clinics. Herein, we developed a novel hybridization chain reaction (HCR)-based electrochemical miRNAs chip (e-miRchip), featured with gold nanostructured electrodes (GNEs) and silver nanoparticle reporters (AgNRs), for sensitive and multiplexed miRNA detection. AgNRs were synthesized and applied on the e-miRchip to generate strong redox signals in the presence of miRNA. The stem–loop capture probe was covalently immobilized on the GNEs, and was opened upon miRNA hybridization to consequently trigger the HCR for signal amplification. The multiple long-repeated DNA helix generated by HCR provides the binding sites for the AgNRs, contributing to the amplification of the electrochemical signals of miRNA hybridization. To optimize the detection sensitivity, GNEs with three distinct structures were electroplated, in which flower-like GNEs were found to be the best electrode morphology for miRNAs analysis. Under optimal conditions, the HCR-based e-miRchip showed an excellent detection performance with an LOD of 0.9 fM and a linear detection range from 1 fM to 10 pM. Moreover, this HCR-based e-miRchip platform was able to effectively distinguish miRNAs from the one- or two-base mismatches. This HCR-based e-miRchip holds great potential as a highly efficient and promising miRNA detection platform for the diagnosis and prognosis of cancer and other diseases in the future.https://www.mdpi.com/2079-4983/16/3/98electrochemical detectionHCRnanostructured electrodemiRNAssilver nanoparticles
spellingShingle Bin Wang
Huiqiang Ma
Mingxing Zhou
Xian Huang
Ying Gan
Hong Yang
Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
Journal of Functional Biomaterials
electrochemical detection
HCR
nanostructured electrode
miRNAs
silver nanoparticles
title Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
title_full Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
title_fullStr Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
title_full_unstemmed Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
title_short Hybridization Chain Reaction-Enhanced Ultrasensitive Electrochemical Analysis of miRNAs with a Silver Nano-Reporter on a Gold Nanostructured Electrode Array
title_sort hybridization chain reaction enhanced ultrasensitive electrochemical analysis of mirnas with a silver nano reporter on a gold nanostructured electrode array
topic electrochemical detection
HCR
nanostructured electrode
miRNAs
silver nanoparticles
url https://www.mdpi.com/2079-4983/16/3/98
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