Development and Optimization of In-house ELISA for Detection of Human IgG Antibody to SARS-CoV-2 Full Length Spike Protein.
Development and Optimization of In-house ELISA for Detection of Human IgG Antibody to SARS-CoV-2 Full Length Spike Protein.
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DOI：
10.3390/pathogens9100803
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发表时间：
2020-09-28
期刊：
影响因子：
--
通讯作者：
Azhar EI
中科院分区：
文献类型：
--
作者：
Alandijany TA;El-Kafrawy SA;Tolah AM;Sohrab SS;Faizo AA;Hassan AM;Alsubhi TL;Othman NA;Azhar EI
由新型严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）引起的新型冠状病毒病（COVID-19）持续大流行，对人类健康构成威胁。尽管如此，许多受影响国家目前正在逐步取消最初为应对大流行而实施的COVID-19限制。所谓的“退出战略”要取得成功，就需要继续监测社区中的病毒传播情况，并评估人群中保护性免疫的普遍程度。血清学测试是实现这些目的的重要工具。本文利用SARS-CoV-2全长刺突(S)重组蛋白，建立并优化了一种间接酶联免疫分析法（ELISA），能够可靠地检测人血清中病毒特异性IgG抗体。重要的是，利用微中和（MN）测定法作为参考试验来评估该测定法的性能。我们开发的ELISA具有100%的灵敏度，98.4%的特异性，98.8%的一致性和高的总体准确性。此外，阳性样品的光密度（OD）值与其MN滴度显著相关。该试验特异性检测针对SARS-CoV-2的人IgG抗体，但不检测针对中东呼吸综合征冠状病毒（MERS-CoV）或人类冠状病毒HKU1 （HCoV-HKU1）的人IgG抗体。这种内部ELISA方案的可用性将对各种诊断和流行病学应用具有价值。
The ongoing coronavirus disease 19 (COVID-19) pandemic, caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), poses a threat to human health. Despite this, many affected countries are now in the process of gradual lifting of COVID-19 restrictions that were initially implemented in response to the pandemic. The success of the so-called “exit strategy” requires continued surveillance of virus circulation in the community and evaluation of the prevalence of protective immunity among population. Serology tests are valuable tools for these purposes. Herein, SARS-CoV-2 full-length spike (S) recombinant protein was utilized to develop and optimize an indirect enzyme-linked immunoassay (ELISA) that enables a reliable detection of virus-specific IgG antibody in human sera. Importantly, the performance of this assay was evaluated utilizing micro-neutralization (MN) assay as a reference test. Our developed ELISA offers 100% sensitivity, 98.4% specificity, 98.8% agreement, and high overall accuracy. Moreover, the optical density (OD) values of positive samples significantly correlated with their MN titers. The assay specifically detects human IgG antibodies directed against SARS-CoV-2, but not those to Middle East respiratory syndrome coronavirus (MERS-CoV) or human coronavirus HKU1 (HCoV-HKU1). The availability of this in-house ELISA protocol would be valuable for various diagnostic and epidemiological applications.
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