论文标题

对人类和动物中SARS-COV-2血清学的简单,敏感和定量的基于FACS的测试

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

论文作者

Ribes, Agnès Maurel, Bessière, Pierre, Guéry, Jean Charles, Featherstone, Eloïse Joly, Bruel, Timothée, Robinot, Remy, Schwartz, Olivier, Volmer, Romain, Abravanel, Florence, Izopet, Jacques, Joly, Etienne

论文摘要

血清学检查对于理解生理病理学和遵循Covid-19大流行的演变非常重要。基于表达SARS-COV-2尖峰蛋白的组织培养细胞流式细胞术(FACS)的测定反复证明,其性能比基于板的基于板的测定法ELISA和CLIA(化学发光免疫 - 测定),并且比侧向流量免疫 - 测定法(LFIA)明显好。在这里,我们描述了一种基于两种Jurkat细胞系的混合物染色的优化且非常简单的FACS分析,表达了高水平的S蛋白(Jurkat-S)或荧光蛋白(Jurkat-R表达M-Cherry或Jurkat-G,表达GFP),表达GFP,以内部阴性控制)。我们表明,Jurkat-S \&R-Flow测试的动态范围要比商业ELISA测试更广泛,并且至少在敏感性和特异性方面表现出色。同样,它比我们最近描述的基于血凝的测试帽更敏感和定量。 Jurkat-Flow测试仅需要几微光的血液。因此,它可用于量化从指刺收集的毛细血管血中的各种Ig同种型。它还可以用来评估小鼠,仓鼠,猫和狗的血清学反应。 FACS测试为想要监测人类或动物的血清学反应的实验室提供了一种非常有吸引力的解决方案,以及这些方法与感染,病毒重新感染的易感性以及对COVID-19的保护如何相关。

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.

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