Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies.
Development and characterization of a quantitative ELISA to detect anti-SARS-CoV-2 spike antibodies.
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DOI：
10.1016/j.heliyon.2021.e08444
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发表时间：
2021-12
期刊：
影响因子：
4
通讯作者：
Mendu DR
中科院分区：
文献类型：
--
作者：
Żak MM;Stock A;Stadlbauer D;Zhang W;Cummings K;Marsiglia W;Zargarov A;Amanat F;Tamayo M;Cordon-Cardo C;Krammer F;Mendu DR
一种新的临床检测和定量的抗体对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的检测方法是改编自内部，基于研究的酶联免疫吸附试验（ELISA）。根据监管指南进行开发和验证，该测试获得了纽约州卫生部（NYSDOH）和食品药品监督管理局（FDA）的紧急使用授权（EUA）。2019年西奈山冠状病毒病（COVID-19）抗体检测是一种正交、定量直接ELISA检测，可检测与新型SARS-CoV-2的受体结合域（RBD）和刺突蛋白反应的抗体。该测定在包被有SARS-CoV-2重组RBD或刺突蛋白的96孔板上进行。该试验分为两个阶段，针对RBD的定性筛选试验和针对全长刺突蛋白的定量试验。该检测使用合并的高滴度血清作为参比标准品。在每次ELISA检测运行中均纳入了阴性的COVID-19前和阳性的COVID-19后PCR确认标本，并在两个不同的地点独立进行检测。西奈山COVID-19血清学的敏感性和特异性分别为92.5%（95%CI：0.785-0.980）和100%（CI：0.939-1.000）。通过在22天内重复单次运行评估运行间精密度;通过在22天内每天重复10次评估运行内精密度。两者均在报告的验收标准范围内（CV ≤ 20%）。这项基于人群的研究揭示了这种新型正交COVID-19血清学检测用于检测和定量SARS-CoV-2抗体的适用性和可靠性，允许广泛的临床应用，包括广泛评估SARS-CoV-2血清阳性率和不同人群亚群的抗体谱。冠状病毒病2019;严重急性呼吸道综合征冠状病毒-2（SARSCoV-2）;抗体检测; COVID-19诊断; ELISA; IgG抗体测定。
A novel clinical assay for the detection and quantitation of antibodies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was adapted from an in-house, research-based enzyme-linked immunosorbent assay (ELISA). Development and validation were performed under regulatory guidelines, and the test obtained emergency use authorization (EUA) from the New York State Department of Health (NYSDOH) and the Food and Drug Administration (FDA). The Mount Sinai coronavirus disease 2019 (COVID-19) antibody assay is an orthogonal, quantitative direct ELISA test which detects antibodies reactive to the receptor binding domain (RBD) and the spike protein of the novel SARS-CoV-2. The assay is performed on 96-well plates coated with either SARS-CoV-2 recombinant RBD or spike proteins. The test is divided into two stages, a qualitative screening assay against RBD and a quantitative assay against the full-length spike protein. The test uses pooled high titer serum as a reference standard. Negative pre-COVID-19 and positive post-COVID-19, PCR-confirmed specimens were incorporated in each ELISA test run, and the assays were performed independently at two different locations. The Mount Sinai COVID-19 serology performed with high sensitivity and specificity, 92.5% (95% CI: 0.785–0.980) and 100% (CI: 0.939–1.000) respectively. Between-run precision was assessed with a single run repeated over 22 days; and within-run precision was assessed with 10 replicates per day over 22 days. Both were within reported acceptance criteria (CV ≤ 20%). This population-based study reveals the applicability and reliability of this novel orthogonal COVID-19 serology test for the detection and quantitation of antibodies against SARS-CoV-2, allowing a broad set of clinical applications, including the broad evaluation of SARS-CoV-2 seroprevalence and antibody profiling in different population subsets. Coronavirus disease 2019; Severe acute respiratory syndrome coronavirus-2 (SARSCoV-2); Antibody tests; Diagnosis of COVID-19; ELISA; IgG antibody assay.
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影响因子：
7.6
作者：
Vu H;Shulenin S;Grolla A;Audet J;He S;Kobinger G;Unfer RC;Warfield KL;Aman MJ;Holtsberg FW
通讯作者：
Holtsberg FW
DOI：
10.4049/jimmunol.2000583
发表时间：
2020-08-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Alsoussi WB;Turner JS;Case JB;Zhao H;Schmitz AJ;Zhou JQ;Chen RE;Lei T;Rizk AA;McIntire KM;Winkler ES;Fox JM;Kafai NM;Thackray LB;Hassan AO;Amanat F;Krammer F;Watson CT;Kleinstein SH;Fremont DH;Diamond MS;Ellebedy AH
通讯作者：
Ellebedy AH
影响因子：
82.9
作者：
Amanat F;Stadlbauer D;Strohmeier S;Nguyen THO;Chromikova V;McMahon M;Jiang K;Arunkumar GA;Jurczyszak D;Polanco J;Bermudez-Gonzalez M;Kleiner G;Aydillo T;Miorin L;Fierer DS;Lugo LA;Kojic EM;Stoever J;Liu STH;Cunningham-Rundles C;Felgner PL;Moran T;García-Sastre A;Caplivski D;Cheng AC;Kedzierska K;Vapalahti O;Hepojoki JM;Simon V;Krammer F
通讯作者：
Krammer F
影响因子：
56.9
作者：
Yu, Jingyou;Tostanoski, Lisa H.;Barouch, Dan H.
通讯作者：
Barouch, Dan H.
影响因子：
--
作者：
Stadlbauer, Daniel;Amanat, Fatima;Krammer, Florian
通讯作者：
Krammer, Florian