Serological assays for severe acute respiratory syndrome coronavirus 2 (BARS-CoV-2), March 2020
Serological assays for severe acute respiratory syndrome coronavirus 2 (BARS-CoV-2), March 2020
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DOI：
10.2807/1560-7917.es.2020.25.16.2000421
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发表时间：
2020-04-23
期刊：
影响因子：
19
通讯作者：
Peiris, Malik
中科院分区：
文献类型：
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作者：
Perera, Ranawaka A. P. M.;Mok, Chris K. P.;Peiris, Malik
背景：持续的冠状病毒病（COVID-19）大流行对卫生系统、经济和社会产生重大影响。评估人群中的感染发病率对于估计疾病严重程度和群体免疫力至关重要，这是校准公共卫生干预措施所必需的。我们以前已经证明，实时实现这一目标以影响公共卫生决策是可能的。目的：我们的目的是开发和评价适用于大规模血清流行病学研究的血清学分析方法。方法：建立ELISA法检测严重急性呼吸综合征冠状病毒2型（SARS-COV-2）刺突蛋白受体结合域（RBD） IgG和IgM抗体。我们结合验证性微中和（MN）和90%斑块减少中和试验（PRNT90）对来自24例病毒学确诊的COVID-19患者的51份血清和来自200名健康对照者的年龄分层血清进行了敏感性和特异性评估。结果：IgG和IgM RBD ELISA、MN和PRNT90在发病29天后呈可靠阳性，在年龄分层的对照组中没有检测到交叉反应。我们发现PRNT90试验在检测抗体方面比传统的组织培养感染剂量激发的MN试验更敏感。肝素化血浆似乎降低了病毒攻击剂量的传染性，并可能混淆中和试验的解释。结论：基于刺突蛋白RBD的IgG ELISA检测SARS-CoV-2抗体，再用PRNT90进行确认，是开展大规模血清流行病学研究的有效方法。
Background: The ongoing coronavirus disease (COVID-19) pandemic has major impacts on health systems, the economy and society. Assessing infection attack rates in the population is critical for estimating disease severity and herd immunity which is needed to calibrate public health interventions. We have previously shown that it is possible to achieve this in real time to impact public health decision making. Aim: Our objective was to develop and evaluate serological assays applicable in large-scale sero-epidemiological studies. Methods: We developed an ELISA to detect IgG and IgM antibodies to the receptor-binding domain (RBD) of the spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). We evaluated its sensitivity and specificity in combination with confirmatory microneutralisation (MN) and 90% plaque reduction neutralisation tests (PRNT90) in 51 sera from 24 patients with virotogically confirmed COVID-19 and in age-stratified sera from 200 healthy controls. Results: IgG and IgM RBD ELISA, MN and PRNT90 were reliably positive after 29 days from illness onset with no detectable cross-reactivity in age-stratified controls. We found that PRNT90 tests were more sensitive in detecting antibody than MN tests carried out with the conventional loo tissue culture infectious dose challenge. Heparinised plasma appeared to reduce the infectivity of the virus challenge dose and may confound interpretation of neutralisation test. Conclusion: Using IgG ELISA based on the RBD of the spike protein to screen sera for SARS-CoV-2 antibody, followed by confirmation using PRNT90, is a valid approach for large-scale sero-epidemiology studies.