DNA Switch: Toehold-Mediated DNA Isothermal Amplification for Dengue Serotyping

SLAS Discov. 2019 Jan;24(1):68-76. doi: 10.1177/2472555218791743. Epub 2018 Jul 31.

Abstract

The inherent ability of nucleic acids to recognize a complementary pair has gained wide popularity in DNA sensor applications. DNA molecules can be produced in bulk and easily incorporated with various nanomaterials for sensing applications. More complex designs and sophisticated DNA sensors have been reported over the years to allow DNA detection in a faster, cheaper, and more convenient manner. Here, we report a DNA sensor designed to function like a switch to turn "on" silver nanocluster (AgNC) generation in the presence of a specific DNA target. By defining the probe region sequence, we are able to tune the color of the AgNC generated in direct relation to the different targets. As a proof of concept, we used dengue RNA-dependent RNA polymerase conserved sequences from all four serotypes as targets. This method was able to distinguish each dengue serotype by generating the serotype-respective AgNCs. The DNA switch was also able to identify and amplify the correct target in a mixture of targets with good specificity. This strategy has a detection limit of between 1.5 and 2.0 µM depending on the sequence of AgNC. The DNA switch approach provides an attractive alternative for single-target or multiplex DNA detection.

Keywords: DNA sensor; dengue; isothermal amplification; silver nanocluster; toehold.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques / methods
  • DNA / genetics*
  • Dengue / genetics*
  • Limit of Detection
  • Metal Nanoparticles / chemistry
  • Nanostructures / chemistry
  • Nucleic Acid Amplification Techniques / methods*
  • RNA-Dependent RNA Polymerase / genetics
  • Sensitivity and Specificity
  • Serotyping / methods*
  • Silver / chemistry

Substances

  • Silver
  • DNA
  • RNA-Dependent RNA Polymerase