A Protein Microarray-Based Respiratory Viral Antigen Testing Platform for COVID-19 Surveillance.

Area of Science:

• Biotechnology
• Infectious Disease Diagnostics
• Molecular Biology

Background:

• The COVID-19 pandemic highlights the need for rapid, high-throughput diagnostic tests for respiratory viruses.
• Co-infections with influenza and common cold viruses complicate diagnosis and treatment.

Purpose of the Study:

• To develop and validate a general workflow for an antibody-based microarray assay.
• To enable multiplexed detection of a respiratory viral panel, including SARS-CoV-2 and influenza.
• To assess the clinical performance of the developed antibody microarray assay.

Main Methods:

• Iterative development including antibody screening and immunofluorescence assay design with signal amplification.
• Assay characterization using recombinant proteins, inactivated viruses, and clinical samples.
• Validation using RT-PCR-confirmed SARS-CoV-2 samples to determine clinical sensitivity and specificity.

Main Results:

• The antibody microarray assay demonstrated 77.2% clinical sensitivity and 100% specificity for SARS-CoV-2 detection.
• Assay performance is comparable to FDA-authorized antigen tests and correlates with RT-PCR cycle threshold (Ct) values.
• The multiplexed assay successfully detected SARS-CoV-2 and influenza virus, differentiating between them.

Conclusions:

• The developed antibody microarray assay is a viable tool for rapid, high-throughput detection of respiratory viruses.
• This protein microarray technology is scalable, automatable, and applicable for both diagnostic and research purposes.
• The assay effectively identifies high viral titers and can distinguish between infections with similar symptoms.