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Disulfide-Induced Self-Assembled Targets: A Novel Strategy for the Label Free Colorimetric Detection of Dnas/Rnas Via Unmodified Gold Nanoparticles Publisher Pubmed



Shokri E1 ; Hosseini M1 ; Davari MD2 ; Ganjali MR3, 4 ; Peppelenbosch MP5 ; Rezaee F5, 6
Authors
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Authors Affiliations
  1. 1. Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
  2. 2. Lehrstuhl fur Biotechnologie, RWTH Aachen University, Aachen, 52056, Germany
  3. 3. Center of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran, Tehran, Iran
  4. 4. Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran
  5. 5. Department of Gastroenterology and Hepatology, Erasmus Medical Center, Rotterdam, Netherlands
  6. 6. Department of Cell Biology, University of Groningen, University Medical Center Groningen, Groningen, Netherlands

Source: Scientific Reports Published:2017


Abstract

A modified non-cross-linking gold-nanoparticles (Au-NPs) aggregation strategy has been developed for the label free colorimetric detection of DNAs/RNAs based on self-assembling target species in the presence of thiolated probes. Two complementary thiol- modified probes, each of which specifically binds at one half of the target introduced SH groups at both ends of dsDNA. Continuous disulfide bond formation at 3′ and 5′ terminals of targets leads to the self-assembly of dsDNAs into the sulfur- rich and flexible products with different lengths. These products have a high affinity for the surface of Au-NPs and efficiently protect the surface from salt induced aggregation. To evaluate the assay efficacy, a small part of the citrus tristeza virus (CTV) genome was targeted, leading to a detection limit of about 5 × 10-9 mol.L-1 over a linear ranged from 20 × 10-9 to 10 × 10-7 mol.L-1. This approach also exhibits good reproducibility and recovery levels in the presence of plant total RNA or human plasma total circulating RNA extracts. Self-assembled targets can be then sensitively distinguished from non-assembled or mismatched targets after gel electrophoresis. The disulfide reaction method and integrating self-assembled DNAs/RNAs targets with bare AuNPs as a sensitive indicator provide us a powerful and simple visual detection tool for a wide range of applications. © The Author(s) 2017.