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Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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 quantified.

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An LC-Immuno-MRM-MS Winterplex Method Development Framework for Respiratory Viral Screening.

Bart Van Puyvelde1, François E Dufrasne2, Sarah Denayer2

  • 1ProGenTomics, Laboratory of Pharmaceutical Biotechnology, Ghent University, 9000 Ghent, Belgium.

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Summary
This summary is machine-generated.

A new assay, Winterplex, enables simultaneous detection of influenza A, influenza B, SARS-CoV-2, and RSV. This mass spectrometry-based method offers a scalable alternative to RT-qPCR for respiratory virus surveillance and pandemic preparedness.

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Area of Science:

  • Clinical diagnostics
  • Virology
  • Mass spectrometry

Background:

  • Reliable large-scale testing for respiratory viruses like influenza, SARS-CoV-2, and RSV is crucial for surveillance and pandemic response.
  • RT-qPCR is the gold standard, but the COVID-19 pandemic revealed the need for scalable, alternative detection platforms.
  • Mass spectrometry (MS)-based methods show promise for multiplex viral detection but require clinical validation.

Purpose of the Study:

  • To develop and validate a novel assay for simultaneous detection of multiple respiratory viruses.
  • To evaluate the performance of the new assay against the current gold standard (RT-qPCR).
  • To highlight the potential of MS-based diagnostics for future pandemic preparedness.

Main Methods:

  • Development of a quantitative immuno-multiple reaction monitoring (iMRM) assay named Winterplex.
  • Simultaneous detection of influenza A, influenza B, RSV, and SARS-CoV-2 using two proteotypic peptides per virus.
  • Validation according to ISO standards for in vitro diagnostics and benchmarking against RT-qPCR.

Main Results:

  • The Winterplex assay demonstrated successful simultaneous detection of the targeted respiratory viruses.
  • The assay's performance was benchmarked against RT-qPCR, indicating its viability as an alternative detection method.
  • Validation according to ISO standards confirmed its suitability for in vitro diagnostics.

Conclusions:

  • The Winterplex assay provides a validated, scalable platform for multiplexed respiratory virus detection.
  • Investment in translating and expanding multiplexed MS-based diagnostic methodologies is essential for enhanced pandemic preparedness.
  • MS-based diagnostics offer flexibility for incorporating diverse targets, enabling rapid responses to emerging viral threats.