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Developing a Paper-Based Antigen Assay to Differentiate between Coronaviruses and SARS-CoV-2 Spike Variants
Delyan Hristov1, Hom Rijal2, Jose Gomez-Marquez3
1Department of Engineering, University of Massachusetts Boston, Boston, Massachusetts 02125, United States.
Analytical Chemistry
|May 26, 2021
Summary
A new paper-based immunoassay can differentiate between SARS-CoV-2 and other coronaviruses, including SARS-CoV-1 and CoV-HKU1, in under 30 minutes. This rapid, flexible diagnostic tool offers a scalable alternative to traditional PCR testing for pandemic surveillance.
Area of Science:
- Biotechnology
- Immunology
- Infectious Disease Diagnostics
Background:
- The COVID-19 pandemic highlights the need for rapid, scalable diagnostic tools beyond traditional methods like RT-PCR.
- RT-PCR, while reliable, is limited by its location dependency, time requirements, and cost.
- Paper-based immunoassays offer a promising alternative for flexible, widely distributable diagnostics.
Purpose of the Study:
- To develop a paper-based immunoassay capable of differentiating between spike proteins of various coronaviruses.
- To assess the assay's ability to distinguish between SARS-CoV-2, SARS-CoV-1, and CoV-HKU1, with minimal cross-reactivity to other human coronaviruses.
- To investigate factors influencing assay performance, such as running media and antigen interference.
Main Methods:
- Development of a novel paper-based immunoassay utilizing distinct binding patterns for antigen identification.
- Testing the assay's specificity against spike proteins from SARS-CoV-1, SARS-CoV-2, CoV-HKU1, HCoV-OC43, and HCoV-229E.
- Evaluation of assay performance using different running media, including human saliva and serum, and assessment of antigen interference.
Main Results:
- The paper-based assay successfully differentiated between SARS-CoV-1, SARS-CoV-2, and CoV-HKU1 spike proteins with negligible cross-reactivity.
- The assay demonstrated the ability to distinguish between fractions of the same spike protein through unique binding patterns at four test line locations.
- Human saliva as a running medium yielded positive results, whereas human serum resulted in the lowest signal quality.
Conclusions:
- A rapid, paper-based immunoassay has been developed for differentiating between specific coronavirus spike proteins.
- This assay offers a flexible, potentially off-the-shelf diagnostic solution for pandemic monitoring, complementing existing methods.
- Further optimization of running media and assay design could enhance its utility for widespread diagnostic applications.
