Infections and Cancer Epidemiology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany.
B Cell Immunology, German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ), 69120 Heidelberg, Germany.
Viruses. 2021 Apr 24;13(5):749. doi: 10.3390/v13050749.
The emerging SARS-CoV-2 pandemic entails an urgent need for specific and sensitive high-throughput serological assays to assess SARS-CoV-2 epidemiology. We, therefore, aimed at developing a fluorescent-bead based SARS-CoV-2 multiplex serology assay for detection of antibody responses to the SARS-CoV-2 proteome. Proteins of the SARS-CoV-2 proteome and protein N of SARS-CoV-1 and common cold Coronaviruses (ccCoVs) were recombinantly expressed in or HEK293 cells. Assay performance was assessed in a COVID-19 case cohort ( = 48 hospitalized patients from Heidelberg) as well as = 85 age- and sex-matched pre-pandemic controls from the ESTHER study. Assay validation included comparison with home-made immunofluorescence and commercial enzyme-linked immunosorbent (ELISA) assays. A sensitivity of 100% (95% CI: 86-100%) was achieved in COVID-19 patients 14 days post symptom onset with dual sero-positivity to SARS-CoV-2 N and the receptor-binding domain of the spike protein. The specificity obtained with this algorithm was 100% (95% CI: 96-100%). Antibody responses to ccCoVs N were abundantly high and did not correlate with those to SARS-CoV-2 N. Inclusion of additional SARS-CoV-2 proteins as well as separate assessment of immunoglobulin (Ig) classes M, A, and G allowed for explorative analyses regarding disease progression and course of antibody response. This newly developed SARS-CoV-2 multiplex serology assay achieved high sensitivity and specificity to determine SARS-CoV-2 sero-positivity. Its high throughput ability allows epidemiologic SARS-CoV-2 research in large population-based studies. Inclusion of additional pathogens into the panel as well as separate assessment of Ig isotypes will furthermore allow addressing research questions beyond SARS-CoV-2 sero-prevalence.
新型严重急性呼吸综合征冠状病毒 2 型（SARS-CoV-2）大流行需要特异性和敏感性高的高通量血清学检测方法来评估 SARS-CoV-2 的流行病学情况。因此，我们旨在开发一种基于荧光珠的 SARS-CoV-2 多重血清学检测方法，用于检测针对 SARS-CoV-2 蛋白质组的抗体反应。SARS-CoV-2 蛋白质组的蛋白质和 SARS-CoV-1 的 N 蛋白以及普通感冒冠状病毒（ccCoV）的 N 蛋白在 或 HEK293 细胞中被重组表达。在海德堡的 48 名住院 COVID-19 患者病例队列和来自 ESTHER 研究的 85 名年龄和性别匹配的大流行前对照中评估了检测方法的性能。检测方法的验证包括与自制免疫荧光和商业酶联免疫吸附（ELISA）检测方法的比较。在 COVID-19 患者出现症状后 14 天，双阳性检测到 SARS-CoV-2 N 和刺突蛋白的受体结合域，获得了 100%（95%CI：86-100%）的敏感性。使用该算法获得的特异性为 100%（95%CI：96-100%）。对 ccCoV N 的抗体反应非常高，与对 SARS-CoV-2 N 的抗体反应无关。包含其他 SARS-CoV-2 蛋白以及单独评估免疫球蛋白（Ig）类别 M、A 和 G 允许针对疾病进展和抗体反应过程进行探索性分析。这种新开发的 SARS-CoV-2 多重血清学检测方法具有高灵敏度和特异性，可用于确定 SARS-CoV-2 血清阳性。其高通量能力允许在大型基于人群的研究中进行 SARS-CoV-2 流行病学研究。将其他病原体纳入面板以及单独评估 Ig 同种型将进一步允许解决 SARS-CoV-2 血清流行率以外的研究问题。
