Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.
Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Centergrid.267301.1, Memphis, Tennessee, USA.
Microbiol Spectr. 2021 Oct 31;9(2):e0105921. doi: 10.1128/Spectrum.01059-21. Epub 2021 Oct 27.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in late 2019 and has since caused a global pandemic resulting in millions of cases and deaths. Diagnostic tools and serological assays are critical for controlling the outbreak, especially assays designed to quantitate neutralizing antibody levels, considered the best correlate of protection. As vaccines become increasingly available, it is important to identify reliable methods for measuring neutralizing antibody responses that correlate with authentic virus neutralization but can be performed outside biosafety level 3 (BSL3) laboratories. While many neutralizing assays using pseudotyped virus have been developed, there have been few studies comparing the different assays to each other as surrogates for authentic virus neutralization. Here, we characterized three enzyme-linked immunosorbent assays (ELISAs) and three pseudotyped vesicular stomatitis virus (VSV) neutralization assays and assessed their concordance with authentic virus neutralization. The most accurate assays for predicting authentic virus neutralization were luciferase- and secreted embryonic alkaline phosphatase (SEAP)-expressing pseudotyped virus neutralizations, followed by green fluorescent protein (GFP)-expressing pseudotyped virus neutralization, and then the ELISAs. The ongoing COVID-19 pandemic is caused by infection with severe acute respiratory syndrome virus 2 (SARS-CoV-2). Prior infection or vaccination can be detected by the presence of antibodies in the blood. Antibodies in the blood are also considered to be protective against future infections from the same virus. The "gold standard" assay for detecting protective antibodies against SARS-CoV-2 is neutralization of authentic SARS-CoV-2 virus. However, this assay can only be performed under highly restrictive biocontainment conditions. We therefore characterized six antibody-detecting assays for their correlation with authentic virus neutralization. The significance of our research is in outlining the advantages and disadvantages of the different assays and identifying the optimal surrogate assay for authentic virus neutralization. This will allow for more accurate assessments of protective immunity against SARS-CoV-2 following infection and vaccination.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）于 2019 年末出现，此后引发了一场全球性大流行，导致数百万人患病和死亡。诊断工具和血清学检测对于控制疫情至关重要，特别是设计用于定量中和抗体水平的检测，被认为是保护的最佳相关指标。随着疫苗的日益普及，确定可靠的方法来测量中和抗体反应非常重要，这些方法与真实病毒中和相关，但可以在生物安全 3 级（BSL3）实验室之外进行。虽然已经开发了许多使用假型病毒的中和检测方法，但很少有研究比较不同的检测方法作为真实病毒中和的替代物。在这里，我们描述了三种酶联免疫吸附测定（ELISA）和三种假型水疱性口炎病毒（VSV）中和测定，并评估了它们与真实病毒中和的一致性。预测真实病毒中和最准确的检测方法是表达荧光素酶和分泌型碱性磷酸酶（SEAP）的假型病毒中和，其次是表达绿色荧光蛋白（GFP）的假型病毒中和，然后是 ELISA。目前的 COVID-19 大流行是由严重急性呼吸综合征病毒 2（SARS-CoV-2）感染引起的。先前的感染或接种疫苗可以通过血液中抗体的存在来检测。血液中的抗体也被认为对来自同一病毒的未来感染具有保护作用。检测针对 SARS-CoV-2 的保护性抗体的“金标准”检测方法是中和真实的 SARS-CoV-2 病毒。然而，这种检测方法只能在高度限制的生物安全条件下进行。因此，我们对六种抗体检测方法进行了特征描述，以确定它们与真实病毒中和的相关性。我们研究的意义在于概述不同检测方法的优缺点，并确定用于真实病毒中和的最佳替代检测方法。这将允许更准确地评估感染和接种疫苗后对 SARS-CoV-2 的保护性免疫。
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