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Nucleic Acid Lateral Flow Assays - A Review on Oligonucleotide-Based Methods.

João M Vindeirinho1,2,3, Eva Pinho1,2,3, Nuno F Azevedo2,3

  • 1INIAV, I.P - National Institute for Agrarian and Veterinarian Research, Vairão, Portugal.

Critical Reviews in Analytical Chemistry
|October 10, 2025
PubMed
Summary
This summary is machine-generated.

Nucleic Acid Lateral Flow Assays (NALFAs) offer sensitive DNA and RNA detection using oligonucleotides. This review details NALFA strategies, performance, and commercial potential, focusing on RNA virus detection.

Keywords:
Molecular diagnosticsnucleic acid detectionnucleic acid lateral flow assaysnucleic acidsoligonucleotides

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Biosensing

Background:

  • Nucleic Acid Lateral Flow Assays (NALFAs) are increasingly vital for DNA and RNA detection, especially for pathogen screening.
  • Traditional NALFAs often depend on antibody-antigen interactions, but oligonucleotide-based recognition offers versatile alternatives.

Purpose of the Study:

  • To review oligonucleotide-based Nucleic Acid Lateral Flow Assays (NALFAs).
  • To cover detection strategies, readout methods, signal amplification, and analytical performance.
  • To highlight commercial applications and emerging innovations in NALFA technology.

Main Methods:

  • Literature review focusing on oligonucleotide-based NALFAs.
  • Analysis of detection and signal amplification strategies.
  • Evaluation of analytical performance and commercial viability.

Main Results:

  • Oligonucleotide-based recognition is a key strategy in modern NALFAs.
  • Various detection and amplification methods enhance NALFA sensitivity and specificity.
  • Significant commercial potential exists, particularly for RNA virus detection, including coronaviruses.

Conclusions:

  • Oligonucleotide-based NALFAs represent a significant advancement in molecular diagnostics.
  • The technology shows strong potential for rapid, specific pathogen detection and commercialization.
  • Emerging innovations promise further improvements in NALFA capabilities.