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Updated: Dec 7, 2025

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection

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High-throughput viral microneutralization method for feline coronavirus using image cytometry.

Morgan Pearson¹, Alora LaVoy¹, Leo Li-Ying Chan²

¹Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, 80523, United States.

Journal of Virological Methods

|September 26, 2020

Summary

A new high-throughput assay using an image cytometer rapidly screens feline coronavirus (FCoV) neutralizing antibodies and antiviral compounds. This method accelerates the development of treatments and vaccines for FCoV infections and other viral diseases.

Keywords:

Celigo Feline coronavirus (FCoV)High-throughput screening (HTS)Image cytometry Plaque-Reduction neutralization tests (PRNT)Viral titer

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Area of Science:

Virology
Immunology
Drug Discovery

Background:

Feline coronaviruses (FCoV) cause asymptomatic enteric infections or fatal feline infectious peritonitis (FIP).
Current methods for assessing FCoV immunity and screening therapeutics, like the plaque reduction neutralization test (PRNT), are slow, labor-intensive, and lack sensitivity.
There is a critical need for efficient assays to develop effective vaccines and antiviral treatments for FCoV.

Purpose of the Study:

To develop and validate a novel, high-throughput viral microneutralization assay for feline coronaviruses (FCoV).
To optimize the assay for rapid screening of therapeutic antibodies, antiviral compounds, and vaccine candidates.
To demonstrate the assay's utility in exploring immune correlates of protection against FCoV.

Main Methods:

A high-throughput viral microneutralization assay was developed using a plate-based image cytometer (Celigo Image Cytometer).
Assay parameters including cell density, surface coating, virus concentration, incubation time, wash buffer, and fluorescent labeling were optimized.
The assay was validated using plasma from naturally FCoV-infected cats to assess immune responses.

Main Results:

The developed assay provides a robust and efficient method for quantifying virus-infected cells.
Optimization resulted in a sensitive and high-throughput platform for neutralization testing.
The assay successfully demonstrated its capability to screen immune responses and potential therapeutics against FCoV.

Conclusions:

The novel high-throughput viral microneutralization assay using the Celigo Image Cytometer is a significant advancement over traditional methods.
This assay enables rapid screening of therapeutic antibodies, antiviral compounds, and vaccines against FCoV.
The platform's efficiency and robustness make it applicable to accelerating the discovery and development of interventions for various viral infectious diseases.