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Sensitive detection of SARS-CoV-2 seroconversion by flow cytometry reveals the presence of nucleoprotein-reactive antibodies in unexposed individuals
medRxiv - Allergy and Immunology Pub Date : 2021-02-18 , DOI: 10.1101/2020.07.28.20162941 Leire Egia-Mendikute , Alexandre Bosch , Endika Prieto-Fernández , So Young Lee , Borja Jiménez-Lasheras , Ana García del Río , Asier Antoñana-Vildosola , Chiara Bruzzone , Maider Bizkarguenaga , Nieves Embade , Rubén Gil-Redondo , María Luz Martínez-Chantar , Marcos López-Hoyos , Nicola G A Abrescia , Jose M. Mato , Oscar Millet , Asís Palazón
medRxiv - Allergy and Immunology Pub Date : 2021-02-18 , DOI: 10.1101/2020.07.28.20162941 Leire Egia-Mendikute , Alexandre Bosch , Endika Prieto-Fernández , So Young Lee , Borja Jiménez-Lasheras , Ana García del Río , Asier Antoñana-Vildosola , Chiara Bruzzone , Maider Bizkarguenaga , Nieves Embade , Rubén Gil-Redondo , María Luz Martínez-Chantar , Marcos López-Hoyos , Nicola G A Abrescia , Jose M. Mato , Oscar Millet , Asís Palazón
There is an ongoing need of developing sensitive and specific methods for the determination of SARS-CoV-2 seroconversion. For this purpose, we have developed a multiplexed flow cytometric bead array (C19BA) that allows the identification of IgG and IgM antibodies against three immunogenic proteins simultaneously: the spike receptor-binding domain (RBD), the spike protein subunit 1 (S1) and the nucleoprotein (N). Using different cohorts of samples collected before and after the pandemic, we show that this assay is more sensitive than ELISAs performed in our laboratory. The combination of three viral antigens allows for the interrogation of full seroconversion. Importantly, we have detected N-reactive antibodies in COVID-19-negative individuals. Here we present an immunoassay that can be easily implemented and has superior potential to detect low antibody titers compared to current gold standard serology methods.
中文翻译:
通过流式细胞仪敏感地检测SARS-CoV-2血清转化,发现未暴露个体中存在核蛋白反应性抗体
持续需要开发灵敏且特定的方法来确定SARS-CoV-2血清转化。为此,我们开发了一种多重流式细胞术微珠阵列(C19BA),可同时鉴定针对三种免疫原性蛋白的IgG和IgM抗体:刺突受体结合域(RBD),刺突蛋白亚基1(S1)和核蛋白(N)。使用大流行之前和之后收集的不同队列样本,我们证明了该检测方法比在我们实验室中进行的ELISA灵敏度更高。三种病毒抗原的组合可进行完全血清转化的询问。重要的是,我们已经在COVID-19阴性个体中检测到N反应性抗体。
更新日期:2021-02-18
中文翻译:
通过流式细胞仪敏感地检测SARS-CoV-2血清转化,发现未暴露个体中存在核蛋白反应性抗体
持续需要开发灵敏且特定的方法来确定SARS-CoV-2血清转化。为此,我们开发了一种多重流式细胞术微珠阵列(C19BA),可同时鉴定针对三种免疫原性蛋白的IgG和IgM抗体:刺突受体结合域(RBD),刺突蛋白亚基1(S1)和核蛋白(N)。使用大流行之前和之后收集的不同队列样本,我们证明了该检测方法比在我们实验室中进行的ELISA灵敏度更高。三种病毒抗原的组合可进行完全血清转化的询问。重要的是,我们已经在COVID-19阴性个体中检测到N反应性抗体。