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一种用于易于实施、具有特异性和定量性的 SARS-CoV-2 血清学检测的全面抗原生产和特性研究。

A comprehensive antigen production and characterisation study for easy-to-implement, specific and quantitative SARS-CoV-2 serotests.

机构信息

Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria.

Department of Biotechnology, University of Natural Resources and Life Sciences (BOKU) Vienna, Muthgasse 18, 1190 Vienna, Austria; Novasign GmbH Vienna, Austria.

出版信息

EBioMedicine. 2021 May;67:103348. doi: 10.1016/j.ebiom.2021.103348. Epub 2021 Apr 25.

DOI:10.1016/j.ebiom.2021.103348
PMID:33906067
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8099623/
Abstract

BACKGROUND

Antibody tests are essential tools to investigate humoral immunity following SARS-CoV-2 infection or vaccination. While first-generation antibody tests have primarily provided qualitative results, accurate seroprevalence studies and tracking of antibody levels over time require highly specific, sensitive and quantitative test setups.

METHODS

We have developed two quantitative, easy-to-implement SARS-CoV-2 antibody tests, based on the spike receptor binding domain and the nucleocapsid protein. Comprehensive evaluation of antigens from several biotechnological platforms enabled the identification of superior antigen designs for reliable serodiagnostic. Cut-off modelling based on unprecedented large and heterogeneous multicentric validation cohorts allowed us to define optimal thresholds for the tests' broad applications in different aspects of clinical use, such as seroprevalence studies and convalescent plasma donor qualification.

FINDINGS

Both developed serotests individually performed similarly-well as fully-automated CE-marked test systems. Our described sensitivity-improved orthogonal test approach assures highest specificity (99.8%); thereby enabling robust serodiagnosis in low-prevalence settings with simple test formats. The inclusion of a calibrator permits accurate quantitative monitoring of antibody concentrations in samples collected at different time points during the acute and convalescent phase of COVID-19 and disclosed antibody level thresholds that correlate well with robust neutralization of authentic SARS-CoV-2 virus.

INTERPRETATION

We demonstrate that antigen source and purity strongly impact serotest performance. Comprehensive biotechnology-assisted selection of antigens and in-depth characterisation of the assays allowed us to overcome limitations of simple ELISA-based antibody test formats based on chromometric reporters, to yield comparable assay performance as fully-automated platforms.

FUNDING

WWTF, Project No. COV20-016; BOKU, LBI/LBG.

摘要

背景

抗体检测是研究 SARS-CoV-2 感染或接种后体液免疫的重要工具。虽然第一代抗体检测主要提供定性结果，但准确的血清流行率研究和随时间推移跟踪抗体水平需要高度特异性、敏感和定量的检测设置。

方法

我们基于刺突受体结合域和核衣壳蛋白开发了两种定量、易于实施的 SARS-CoV-2 抗体检测方法。来自多个生物技术平台的抗原的综合评估使我们能够识别出优越的抗原设计，用于可靠的血清诊断。基于前所未有的大型和异质多中心验证队列的截止值建模允许我们为测试的广泛应用定义最佳阈值，例如血清流行率研究和恢复期血浆供体资格认证。

发现

两种开发的血清学检测各自与全自动 CE 标记测试系统一样表现出色。我们描述的灵敏度改进正交测试方法确保了最高的特异性（99.8%）；从而能够在低流行率环境中使用简单的测试格式进行稳健的血清诊断。包含校准器可准确监测 COVID-19 急性期和恢复期采集的样本中抗体浓度的定量监测，并揭示与真实 SARS-CoV-2 病毒的强大中和作用相关的抗体水平阈值。

解释

我们证明了抗原来源和纯度强烈影响血清学检测性能。对抗原的综合生物技术辅助选择和对检测的深入表征使我们能够克服基于比色报告器的简单 ELISA 抗体检测格式的限制，从而获得与全自动平台相当的检测性能。

资助

WWTF，项目编号 COV20-016；BOKU，LBI/LBG。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/5abfad6dd25f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/09e51bdd3ba0/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/3fb97af1570b/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/a832546710c9/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/6e80e1bdd7c7/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/5abfad6dd25f/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/09e51bdd3ba0/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/3fb97af1570b/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/a832546710c9/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/6e80e1bdd7c7/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9c49/8099623/5abfad6dd25f/gr5.jpg

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