A label-free impedimetric genosensor for the nucleic acid amplification-free detection of extracted RNA of dengue virus

Ching Chou Wu, Hao Yu Yen, Lu Ting Lai, Guey Chuen Perng, Cheng Rei Lee, Shuenn Jue Wu

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

Developing rapid and sensitive diagnostic methods for dengue virus (DENV) infection is of prime priority because DENV infection is the most prevalent mosquito-borne viral disease. This work proposes an electrochemical impedance spectroscopy (EIS)-based genosensor for the label-free and nucleic acid amplification-free detection of extracted DENV RNA intended for a sensitive diagnosis of DENV infection. A concentration ratio of 0.04 mM 6-mercaptohexanoic acid (MHA) to 1 mM 6-mercapto-1-hexanol (MCH) was selected to modify thin-film gold electrodes as a link to control the coverage of self-designed probe DNA (pDNA) at a density of 4.5 ± 0.4 × 1011 pDNA/cm2. The pDNA/MHA/MCH-modified genosensors are proven to improve the hybridization efficiency of a synthetic 160-mer target DNA (160mtDNA) with a 140-mer electrode side overhang as compared to other MHA/MCH ratio-modified genosensors. The MHA(0.04 mM)/MCH(1 mM)-modified genosensors also present good hybridization efficiency with the extracted DENV serotype 1 (DENV1) RNA samples, having the same electrode side overhangs with the 160mtDNA, showing a low detection limit of 20 plaque forming units (PFU)/mL, a linear range of 102–105 PFU/mL and good selectivity for DENV1. The pDNA density-controlled method has great promise to construct sensitive genosensors based on the hybridization of extracted DENV nucleic acids.

Original languageEnglish
Article number3728
Pages (from-to)1-15
Number of pages15
JournalSensors (Switzerland)
Volume20
Issue number13
DOIs
Publication statusPublished - 2020 Jul 1

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Information Systems
  • Biochemistry
  • Atomic and Molecular Physics, and Optics
  • Instrumentation
  • Electrical and Electronic Engineering

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