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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Related Experiment Video

Updated: Dec 2, 2025

A Rapid, Multiplex Dual Reporter IgG and IgM SARS-CoV-2 Neutralization Assay for a Multiplexed Bead-Based Flow Analysis System
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A Multiplex Microsphere IgG Assay for SARS-CoV-2 Using ACE2-Mediated Inhibition as a Surrogate for Neutralization.

Andrew Cameron1, Claire A Porterfield2, Larry D Byron1

  • 1University of Rochester Medical Center, Clinical Microbiology, Department of Pathology and Laboratory Medicine, Rochester, New York, USA.

Journal of Clinical Microbiology
|November 3, 2020
PubMed
Summary
This summary is machine-generated.

A new 3Flex assay detects antibodies to SARS-CoV-2 antigens with high sensitivity and specificity. This flexible, high-throughput platform aids in diagnosing COVID-19 and understanding immune responses to the virus.

Keywords:
COVID-19IgGSARS-CoV-2coronavirusfluorescence assaysimmunoassaysmicrosphereneutralizing antibodiesserology

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Area of Science:

  • Immunology
  • Virology
  • Medical Diagnostics

Background:

  • The COVID-19 pandemic necessitates reliable serological tests for diagnosing SARS-CoV-2 infection and for epidemiological surveillance.
  • Existing serological assays face challenges in throughput, sensitivity, and specificity, particularly for novel pathogens.
  • Accurate antibody detection is crucial for understanding disease progression and population immunity.

Purpose of the Study:

  • To develop and evaluate a multiplex fluorescent microsphere-based assay, named 3Flex, for detecting antibodies against key SARS-CoV-2 antigens.
  • To assess the sensitivity and specificity of the 3Flex assay compared to a commercially available assay.
  • To characterize the kinetics of antibody responses to SARS-CoV-2 infection.

Main Methods:

  • Developed a 3Flex assay detecting antibodies to SARS-CoV-2 spike (S) protein, receptor-binding domain (RBD), and nucleocapsid (NP) antigens.
  • Assessed specificity using 213 prepandemic serum samples.
  • Measured sensitivity using serum from 125 COVID-19 patients at different time points post-symptom onset and compared it with the Abbott Architect SARS-CoV-2 IgG assay.
  • Analyzed antibody response kinetics in 140 COVID-19 patients using 534 serum samples.

Main Results:

  • The 3Flex assay demonstrated high specificity using prepandemic samples.
  • Early in infection (≤5 days from symptom onset), 3Flex showed higher sensitivity (48.0%) than the Abbott assay (32.0%).
  • Both assays performed comparably for samples collected ≥21 days post-symptom onset.
  • Characterized the rise, peak, and decline of antibody responses to S, RBD, and NP antigens, noting significant inter-individual variation.
  • Demonstrated inhibition of antibody binding to S and RBD by soluble ACE2.

Conclusions:

  • The 3Flex assay is a sensitive and specific tool for SARS-CoV-2 antibody detection.
  • The assay's flexible, high-throughput nature makes it adaptable for emerging infectious diseases.
  • The findings provide insights into the dynamics of antibody responses following SARS-CoV-2 infection.
  • This platform can support both diagnostic applications and large-scale serological surveillance efforts.