Highly Sensitive and Specific Multiplex Antibody Assays To Quantify Immunoglobulins M, A, and G against SARS-CoV-2 Antigens.
Highly Sensitive and Specific Multiplex Antibody Assays To Quantify Immunoglobulins M, A, and G against SARS-CoV-2 Antigens.
复制标题
高度敏感和特异性的多重抗体测定法对SARS-COV-2抗原量化免疫球蛋白M，A和G。
DOI：
10.1128/jcm.01731-20
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发表时间：
2021-01-21
影响因子：
9.4
通讯作者：
Moncunill G
中科院分区：
文献类型：
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作者：
Dobaño C;Vidal M;Santano R;Jiménez A;Chi J;Barrios D;Ruiz-Olalla G;Rodrigo Melero N;Carolis C;Parras D;Serra P;Martínez de Aguirre P;Carmona-Torre F;Reina G;Santamaria P;Mayor A;García-Basteiro AL;Izquierdo L;Aguilar R;Moncunill G
需要可靠的血清学检测来确定严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）抗体的流行率，并表征对该疾病的免疫力，以解决2019冠状病毒病（COVID-19）大流行的关键知识缺口。基于xMAP Luminex平台的定量悬浮阵列技术（qSAT）检测克服了快速诊断检测和酶联免疫吸附测定（ELISA）的局限性，具有更高的精度、动态范围、通量、小型化、成本效益和多路复用能力。需要可靠的血清学检测来确定严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）抗体的流行率，并表征对该疾病的免疫力，以解决2019冠状病毒病（COVID-19）大流行的关键知识缺口。基于xMAP Luminex平台的定量悬浮阵列技术（qSAT）检测克服了快速诊断检测和酶联免疫吸附测定（ELISA）的局限性，具有更高的精度、动态范围、通量、小型化、成本效益和多路复用能力。我们开发了三种针对一组八种SARS-CoV-2抗原的IgM、伊加和IgG的qSAT检测方法，包括刺突蛋白（S）、核衣壳蛋白（N）和膜蛋白（M）构建体。对测定进行了优化，以最大限度地减少处理时间并最大限度地提高信噪比。我们使用128份大流行前血浆样本（阴性对照）和104份来自SARS-CoV-2诊断个体的血浆样本（阳性对照）评估了他们的表现，其中5人无症状，51人有轻度症状，48人住院。在阴性对照中检测到识别N、M和S蛋白的既存IgG抗体，这表明与普通感冒冠状病毒具有交叉反应性。表现最好的抗体/抗原特征在症状发作后≥14天和≥21天的特异性为100%，灵敏度为95.78%和95.65%，曲线下面积（AUC）分别为0.977和0.999。通过qSAT测定法评估的组合多种标志物具有最高的效率、广度和通用性，可准确检测低水平抗体应答，以获得关于人群中暴露于新型病原体的患病率的可靠数据。我们的检测将有助于深入了解抗体与免疫力及其动力学的相关性，这是疫苗开发以抗击COVID-19大流行所需的。
Reliable serological tests are required to determine the prevalence of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and to characterize immunity to the disease in order to address key knowledge gaps in the coronavirus disease 2019 (COVID-19) pandemic. Quantitative suspension array technology (qSAT) assays based on the xMAP Luminex platform overcome the limitations of rapid diagnostic tests and enzyme-linked immunosorbent assays (ELISAs) with their higher precision, dynamic range, throughput, miniaturization, cost-efficiency, and multiplexing capacity. Reliable serological tests are required to determine the prevalence of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and to characterize immunity to the disease in order to address key knowledge gaps in the coronavirus disease 2019 (COVID-19) pandemic. Quantitative suspension array technology (qSAT) assays based on the xMAP Luminex platform overcome the limitations of rapid diagnostic tests and enzyme-linked immunosorbent assays (ELISAs) with their higher precision, dynamic range, throughput, miniaturization, cost-efficiency, and multiplexing capacity. We developed three qSAT assays for IgM, IgA, and IgG against a panel of eight SARS-CoV-2 antigens, including spike protein (S), nucleocapsid protein (N), and membrane protein (M) constructs. The assays were optimized to minimize the processing time and maximize the signal-to-noise ratio. We evaluated their performances using 128 prepandemic plasma samples (negative controls) and 104 plasma samples from individuals with SARS-CoV-2 diagnosis (positive controls), of whom 5 were asymptomatic, 51 had mild symptoms, and 48 were hospitalized. Preexisting IgG antibodies recognizing N, M, and S proteins were detected in negative controls, which is suggestive of cross-reactivity to common-cold coronaviruses. The best-performing antibody/antigen signatures had specificities of 100% and sensitivities of 95.78% at ≥14 days and 95.65% at ≥21 days since the onset of symptoms, with areas under the curve (AUCs) of 0.977 and 0.999, respectively. Combining multiple markers as assessed by qSAT assays has the highest efficiency, breadth, and versatility to accurately detect low-level antibody responses for obtaining reliable data on the prevalence of exposure to novel pathogens in a population. Our assays will allow gaining insights into antibody correlates of immunity and their kinetics, required for vaccine development to combat the COVID-19 pandemic.