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SPR-based single nucleotide mismatch biosensor.

Eftim Milkani1,2, Aung M Khaing2,3, Sergi Morais4

  • 1Chemistry and Biochemistry, Worcester Polytechnic Institute, 100 Institute Road, Worcester, MA 01609, USA.

Analytical Methods : Advancing Methods and Applications
|September 17, 2020
PubMed
Summary
This summary is machine-generated.

This study presents a novel DNA sensor using electrochemical impedance spectroscopy and surface plasmon resonance (SPR) for detecting DNA and RNA hybridization. The sensor achieves a 20 pM detection limit and demonstrates reusability over 20 cycles.

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Area of Science:

  • Biosensing
  • Nanotechnology
  • Surface Chemistry

Background:

  • DNA hybridization detection is crucial for diagnostics.
  • Developing sensitive and reusable biosensors is an ongoing challenge.
  • Surface immobilization techniques are key for sensor performance.

Purpose of the Study:

  • To develop and characterize a DNA biosensor for detecting unlabeled DNA oligonucleotides.
  • To evaluate the sensor's performance using electrochemical impedance spectroscopy and surface plasmon resonance (SPR).
  • To assess the sensor's specificity, reusability, and operational stability.

Main Methods:

  • Immobilization of thiol-modified DNA probes onto a gold surface via self-assembly.
  • Detection of DNA and RNA hybridization using electrochemical impedance spectroscopy and SPR.
  • Characterization of sensor response to full match, single mismatch, and distal end mismatch DNA targets.

Main Results:

  • A detection limit of 20 pM was achieved for full match target DNA hybridization.
  • The sensor successfully detected RNA hybridization with a similar detection limit.
  • SPR distinguished between full match and some mismatch sequences, but not distal end mismatches.
  • The SPR sensor demonstrated reusability over at least 20 cycles and was insensitive to flow rate and temperature variations.

Conclusions:

  • The developed SPR-based DNA sensor offers sensitive and specific detection of DNA and RNA hybridization.
  • The sensor's reusability and robustness make it a promising platform for various applications.
  • Further optimization may be needed to improve discrimination against all types of mismatches.