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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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Capillary flow-driven immunoassay platform for COVID-19 antigen diagnostics.

Jeremy S Link¹, Cody S Carrell¹, Ilhoon Jang²

¹Department of Chemistry, Colorado State University, USA.

Analytica Chimica Acta

|August 21, 2023

Summary

A new Capillary-Driven Immunoassay (CaDI) device offers rapid, affordable SARS-CoV-2 diagnostics. This simple, visual test provides results comparable to lab-based methods, enhancing point-of-care testing capabilities.

Keywords:

COVID-19 Capillary-driven microfluidics Diagnostic testing Immunoassays Point-of-care SARS-CoV-2

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Published on: June 23, 2022

Area of Science:

Biomedical Engineering
Diagnostic Technologies
Microfluidics

Background:

The COVID-19 pandemic highlighted the need for rapid, affordable diagnostic tests.
Traditional methods like PCR and ELISA are sensitive but costly and time-consuming.
Existing point-of-care lateral flow assays (LFAs) often lack sensitivity compared to lab methods.

Purpose of the Study:

To develop and validate a novel Capillary-Driven Immunoassay (CaDI) device.
To automate ELISA steps using microfluidics and capillary action for visual readout.
To achieve rapid, sensitive, and affordable SARS-CoV-2 antigen detection.

Main Methods:

The CaDI device utilizes microfluidic channels and capillary action for passive automation of ELISA steps.
Colorimetric detection is employed, with sequential delivery of reagents, washes, and substrate to a nitrocellulose strip.
SARS-CoV-2 antigen detection was validated using both naked-eye visual assessment and smartphone-based image analysis (ImageJ).

Main Results:

The CaDI device detected SARS-CoV-2 antigen visually in 15-20 minutes.
Analytical detection limits were 83 PFU/mL in buffer and 222 PFU/mL in spiked nasal swabs.
Visual detection limit was determined to be 100 PFU/mL in contrived nasal swabs by untrained users.

Conclusions:

The CaDI device provides a simple, rapid, and visually interpretable method for SARS-CoV-2 antigen detection.
Its performance is comparable to traditional well-plate ELISA methods.
The CaDI platform is adaptable for various immunoassay applications beyond SARS-CoV-2 detection.