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Quantitative circular flow immunoassays with trained object recognition to detect antibodies to SARS-CoV-2 membrane
Ryan Yuki Huang1, Deron Raymond Herr2
1Canyon Crest Academy, San Diego, USA.
Insights
A new multiplexed circular flow immunoassay (CFIA) test strip can simultaneously quantify antibodies for multiple pathogens like SARS-CoV-2 and H1N1. This digital tool offers a practical solution for monitoring immune responses during infectious disease outbreaks.
Area of Science:
- Immunology
- Biotechnology
- Infectious Diseases
Background:
- Effective infectious disease control requires tools to quantitatively monitor pathogen-specific antibodies.
- Current serological lateral flow immunoassays (LFIA) are limited to detecting antibodies for a single antigen.
- There is a need for advanced diagnostic platforms enabling simultaneous detection and quantification of antibodies against multiple pathogens.
Purpose of the Study:
- To develop and validate a multiplexed circular flow immunoassay (CFIA) test strip for simultaneous antibody quantification.
- To integrate YOLO v4 object recognition for automated analysis of CFIA results.
- To assess the CFIA's capability in differentiating and quantifying antibodies against SARS-CoV-2 and H1N1 influenza virus.
Main Methods:
- Fabrication of a multiplexed CFIA test strip capable of detecting multiple antibody targets.
- Utilized YOLO v4-based object recognition for automated image analysis and quantification of spot intensities.
- Quantified antibody titers by measuring spot intensities relative to immunoglobulin G (IgG) reaction spots to correct for image heterogeneity.
- Captured real-time images of the CFIA using an integrated camera for immediate data display.
Main Results:
- The CFIA successfully quantified and differentiated antibodies against SARS-CoV-2 (membrane glycoprotein) and H1N1 (hemagglutinin) in immunized mouse sera.
- Spot intensities correlated with antibody titers and were accurately quantified relative to IgG controls.
- The system demonstrated real-time data display, integrating quantitative results with captured images.
Conclusions:
- CFIA is a novel, specific, and quantitative tool for multi-pathogen antibody detection.
- The developed CFIA platform holds significant potential for digital and simultaneous monitoring of immune responses to various infectious agents.
- This technology offers a practical advancement for disease surveillance and outbreak management.
Abstract:
Amidst infectious disease outbreaks, a practical tool that can quantitatively monitor individuals' antibodies to pathogens is vital for disease control. The currently used serological lateral flow immunoassays (LFIAs) can only detect the presence of antibodies for a single antigen. Here, we fabricated a multiplexed circular flow immunoassay (CFIA) test strip with YOLO v4-based object recognition that can quickly quantify and differentiate antibodies that bind membrane glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) or hemagglutinin of influenza A (H1N1) virus in the sera of immunized mice in one assay using one sample. Spot intensities were found to be indicative of antibody titers to membrane glycoprotein of SARS-CoV-2 and were, thus, quantified relative to spots from immunoglobulin G (IgG) reaction in a CFIA to account for image heterogeneity. Quantitative intensities can be displayed in real time alongside an image of CFIA that was captured by a built-in camera. We demonstrate for the first time that CFIA is a specific, multi-target, and quantitative tool that holds potential for digital and simultaneous monitoring of antibodies recognizing various pathogens including SARS-CoV-2.
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