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Semiquantitative Paper-Based Microfluidic Surrogate Virus Neutralization Test for SARS-CoV-2 Neutralizing Antibodies.

Md Almostasim Mahmud1,2,3,4, Liangcheng Henry Xu1, Anat Usatinsky1

  • 1Department of Chemistry and Biology, Toronto Metropolitan University, Toronto, Ontario M5B 2K3, Canada.

Analytical Chemistry
|July 9, 2024
PubMed
Summary
This summary is machine-generated.

A new paper-based lateral flow assay (LFA) device enables rapid, instrument-free detection of SARS-CoV-2 neutralizing antibodies (nAbs). This surrogate virus neutralization test (sVNT) simplifies immunity measurement without high biosafety levels.

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Area of Science:

  • Immunology
  • Biotechnology
  • Medical Diagnostics

Background:

  • Neutralizing antibodies (nAbs) are crucial for immunity following infection or vaccination.
  • Measuring virus-specific nAb titers is vital for assessing acquired immunity.
  • Traditional methods for nAb titration require high biosafety levels, limiting accessibility.

Purpose of the Study:

  • To develop a paper-based lateral flow assay (LFA) for semiquantitative determination of SARS-CoV-2 neutralizing antibodies (nAbs).
  • To create a rapid, instrument-free diagnostic tool for assessing immunity.
  • To adapt the surrogate virus neutralization test (sVNT) for a user-friendly format.

Main Methods:

  • Fabrication of a paper-based LFA device utilizing a gradient assay design.
  • Implementation of the surrogate virus neutralization test (sVNT) with spike glycoprotein receptor binding domain (RBD) and ACE2 as proxies.
  • Use of gold nanoparticle-conjugated RBD and increasing concentrations of ACE2 for colorimetric readout.
  • Testing with standard nAb solutions and human serum samples.

Main Results:

  • The LFA device successfully provided semiquantitative results for SARS-CoV-2 nAbs.
  • The number of developed colorimetric spots was inversely proportional to nAb concentration.
  • Direct, instrument-free colorimetric detection was achieved.
  • The assay demonstrated effectiveness with both standard solutions and clinical samples.

Conclusions:

  • The developed paper-based LFA is a viable tool for semiquantitative assessment of SARS-CoV-2 nAbs.
  • This sVNT-based LFA offers a simplified, low-biohazard alternative to traditional methods.
  • The device facilitates rapid and accessible measurement of virus-specific neutralizing antibody levels.