Thomas Archana, Messer William B, Hansel Donna E, Streblow Daniel N, Kazmierczak Steven C, Lyski Zoe L, Lu Zhengchun, Slifka Mark K
Division of Neuroscience, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA.
Department of Molecular Microbiology and Immunology, Program in Epidemiology, OHSU-PSU School of Public Health, Portland, Oregon, USA.
Open Forum Infect Dis. 2021 Feb 2;8(3):ofab061. doi: 10.1093/ofid/ofab061. eCollection 2021 Mar.
Serological confirmation of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for understanding the dynamics of the pandemic and determining seroprevalence rates within afflicted communities. Common challenges with SARS-CoV-2 serological assays include poor analytical specificity and sensitivity and lack of a serological standard for quantitative assessment of antibody titers.
To overcome these obstacles, we developed a quantitative enzyme-linked immunosorbent assay based on an optimized 2-dimensional screening assay that utilizes SARS-CoV-2 receptor binding domain (RBD) of spike protein and SARS-CoV-2 spike S1 subunit.
A total of 4 SARS-CoV-2-reactive monoclonal antibodies were evaluated for use as serum standards for calibrating assays performed on different days or by different laboratories. This approach provided quantitative analysis of hospitalized reverse transcription polymerase chain reaction-confirmed COVID-19 cases that in some cases reached >100 μg/mL. The assay demonstrated 72% sensitivity based on time points ranging from 2 to 52 days post-symptom onset, with 100% sensitivity at time points measured ≥13 days post-symptom onset and 100% specificity.
Using these optimized reagents and serological standards, we believe this approach will be useful for sensitive and specific determination of seroconversion rates and quantitatively measuring the durability of antiviral antibody responses following SARS-CoV-2 infection or vaccination.
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的2019冠状病毒病（COVID-19）的血清学确认对于了解大流行动态和确定受灾社区的血清阳性率至关重要。SARS-CoV-2血清学检测的常见挑战包括分析特异性和敏感性差，以及缺乏用于定量评估抗体滴度的血清学标准。
为克服这些障碍，我们基于优化的二维筛选检测方法开发了一种定量酶联免疫吸附测定，该方法利用刺突蛋白的SARS-CoV-2受体结合域（RBD）和SARS-CoV-2刺突S1亚基。
共评估了4种与SARS-CoV-2反应的单克隆抗体，用作校准在不同日期或由不同实验室进行的检测的血清标准品。这种方法对住院的经逆转录聚合酶链反应确诊的COVID-19病例进行了定量分析，在某些情况下抗体水平超过100μg/mL。该检测方法在症状出现后2至52天的时间点显示出72%的敏感性，在症状出现后≥13天测量的时间点敏感性为100%，特异性为100%。
使用这些优化的试剂和血清学标准品，我们相信这种方法将有助于灵敏且特异地测定血清转化率，并定量测量SARS-CoV-2感染或疫苗接种后抗病毒抗体反应的持久性。
