Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

  • Jun Xu
  • Qing Tang
  • Runhui Zhang
  • Haoyi Chen
  • Xinguo Zhang
  • Yue Chen
  • Hong Yan
  • Jincheng Li
  • Huaze Shao
  • Lihong Liu

Related Research Unit(s)

Detail(s)

Original languageEnglish
Journal / PublicationJournal of Pharmaceutical Analysis
Online published1 Jun 2022
Publication statusOnline published - 1 Jun 2022

Abstract

The identification of tumor-related microRNAs (miRNAs) exhibits excellent promise for the early diagnosis of cancer and other bioanalytical applications. Therefore, we developed a sensitive and efficient biosensor using polyadenine (polyA)-mediated fluorescent spherical nucleic acid (FSNA) for miRNA analysis based on strand displacement reactions on gold nanoparticle (AuNP) surfaces and electrokinetic signal amplification (ESA) on a microfluidic chip. In this FSNA, polyA-DNA biosensor was anchored on AuNP surfaces via intrinsic affinity between adenine and Au. The upright conformational polyA-DNA recognition block hybridized with 6-carboxyfluorescein-labeled reporter-DNA, resulting in fluorescence quenching of FSNA probes induced by AuNP-based resonance energy transfer. Reporter DNA was replaced in the presence of target miRNA, leading to the recovery of reporter-DNA fluorescence. Subsequently, reporter-DNAs were accumulated and detected in the front of with Nafion membrane in the microchannel by ESA. Our method showed high selectivity and sensitivity with a limit of detection of 1.3 pM. This method could also be used to detect miRNA-21 in human serum and urine samples, with recoveries of 104.0%–113.3% and 104.9%–108.0%, respectively. Furthermore, we constructed a chip with three parallel channels for the simultaneous detection of multiple tumor-related miRNAs (miRNA-21, miRNA-141, and miRNA-375), which increased the detection efficiency. Our universal method can be applied to other DNA/RNA analyses by altering recognition sequences.

Research Area(s)

  • MicroRNAs, microfluidic chip, electrokinetic signal amplification, polyadenine-DNA, gold nanoparticle

Citation Format(s)

Sensitive detection of microRNAs using polyadenine-mediated fluorescent spherical nucleic acids and a microfluidic electrokinetic signal amplification chip. / Xu, Jun; Tang, Qing; Zhang, Runhui; Chen, Haoyi; Khoo, Bee Luan; Zhang, Xinguo; Chen, Yue; Yan, Hong; Li, Jincheng; Shao, Huaze; Liu, Lihong.

In: Journal of Pharmaceutical Analysis, 01.06.2022.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review