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Quantitative SERS-Based Sandwich-Hybridization Assay for Nucleic Acid Detection.

Kosar Shahsavar1,2, Amr Mostafa1, Dina Mahdi-Joest1

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This study introduces a simple Surface-Enhanced Raman Spectroscopy (SERS) assay for detecting SARS-CoV-2 DNA. The method uses readily available materials for sensitive viral detection and quantification.

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Area of Science:

  • Biochemistry
  • Nanotechnology
  • Spectroscopy

Background:

  • Surface-enhanced Raman spectroscopy (SERS) shows promise for biological molecule quantification, especially DNA.
  • Developing straightforward assays for detecting viral DNA like SARS-CoV-2 is crucial for diagnostics.

Purpose of the Study:

  • To develop a simple, SERS-based sandwich-hybridization assay for detecting SARS-CoV-2 DNA.
  • To utilize commercially available components and standard procedures, avoiding complex synthesis.

Main Methods:

  • The assay uses gold nanoparticle-conjugated TAMRA-labeled DNA nanoprobes and biotin-labeled surface DNA strands.
  • A sandwich structure forms upon target analyte hybridization and is captured on a neutravidin-coated glass via avidin-biotin interaction.
  • Raman signals from the TAMRA dye are enhanced by resonance and plasmonic effects for sensitive detection.

Main Results:

  • The assay successfully detected target DNA, with quantification achieved through spectral analysis and binary image-based analysis.
  • The binary image-based analysis proved reliable for evaluating assay performance.
  • Atomic Force Microscopy (AFM) characterization showed strong correlation with SERS results, validating the approach.

Conclusions:

  • The developed SERS assay offers a sensitive and reliable method for detecting viral DNA.
  • The assay's simplicity and use of standard components make it accessible for widespread application.
  • The combination of SERS and AFM provides accurate and trustworthy results for DNA quantification.