A simple, sensitive and quantitative FACS-based test for SARS-CoV-2 serology in humans and animals

TL;DR

This paper introduces a simple, sensitive, and quantitative FACS-based serology test for SARS-CoV-2 that works well in humans and animals, requiring minimal blood and offering broad dynamic range and high sensitivity.

Contribution

The authors developed an optimized FACS assay using Jurkat cell lines that improves sensitivity, quantification, and versatility over existing serological tests for SARS-CoV-2.

Findings

Broader dynamic range than commercial ELISA

Comparable or better sensitivity and specificity

Effective in small blood samples from finger pricks

Abstract

Serological tests are important for understanding the physiopathology and following the evolution of the Covid-19 pandemic. Assays based on flow cytometry (FACS) of tissue culture cells expressing the spike (S) protein of SARS-CoV-2 have repeatedly proven to perform slightly better than the plate-based assays ELISA and CLIA (chemiluminescent immuno-assay), and markedly better than lateral flow immuno-assays (LFIA). Here, we describe an optimized and very simple FACS assay based on staining a mix of two Jurkat cell lines, expressing either high levels of the S protein (Jurkat-S) or a fluorescent protein (Jurkat-R expressing m-Cherry, or Jurkat-G, expressing GFP, which serve as an internal negative control). We show that the Jurkat-S\&R-flow test has a much broader dynamic range than a commercial ELISA test and performs at least as well in terms of sensitivity and specificity. Also, it is more sensitive and quantitative than the hemagglutination-based test HAT, which we described recently. The Jurkat-flow test requires only a few microliters of blood; thus, it can be used to quantify various Ig isotypes in capillary blood collected from a finger prick. It can be used also to evaluate serological responses in mice, hamsters, cats and dogs. FACS tests offer a very attractive solution for laboratories with access to tissue culture and flow cytometry who want to monitor serological responses in humans or in animals, and how these relate to susceptibility to infection, or re-infection, by the virus, and to protection against Covid-19.