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Ultra-Sensitive 3D Lateral Flow Assay Device for SARS-CoV-2 Detection Based on One-Step Dual-Signal Amplification.

Yi-Ru Chiou1, Wei Wang1, Yuh-Shiuan Chien1

  • 1Institute of Applied Mechanics, National Taiwan University, Taipei 106, Taiwan.

Analytical Chemistry
|November 19, 2025
PubMed
Summary
This summary is machine-generated.

A novel nucleic acid lateral flow assay (NALFA) enables rapid, on-site detection of SARS-CoV-2. This assay uses dual-signal amplification for enhanced sensitivity, providing visual results in 25 minutes.

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Area of Science:

  • Biotechnology
  • Nanotechnology
  • Assay Development

Background:

  • Accurate and rapid detection of SARS-CoV-2 is crucial for pandemic control.
  • Existing diagnostic methods may require complex laboratory procedures or lack on-site applicability.
  • Need for sensitive and user-friendly diagnostic tools for resource-limited settings.

Purpose of the Study:

  • To develop a one-step nucleic acid lateral flow assay (NALFA) for rapid, on-site detection of SARS-CoV-2.
  • To enhance signal amplification for improved sensitivity and colorimetric detection.
  • To create a stable and interference-free assay platform for visual results.

Main Methods:

  • Developed a NALFA platform utilizing polyadenine (polyA) for gold nanoparticle (AuNP) probe functionalization.
  • Implemented bimetallic deposition (Au and Ag staining) for dual-signal amplification.
  • Constructed three-dimensional paper-based flow channels with pre-dried reagents for enhanced stability and reduced interference.

Main Results:

  • Achieved a limit of detection of 2.24 nM for SARS-CoV-2.
  • Demonstrated 100-fold higher signal amplification compared to conventional lateral flow assays (LFAs).
  • Successfully detected SARS-CoV-2 Omicron BA.1 variant with visual results obtained in 25 minutes.

Conclusions:

  • The developed NALFA platform offers a sensitive, rapid, and visual method for SARS-CoV-2 detection.
  • The assay's design with dual-signal amplification and pre-dried reagents enhances performance and stability.
  • This technology holds significant potential for point-of-care diagnostics, especially in resource-limited areas.