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 E. coli or HEK293 cells. Assay performance was assessed in a COVID-19 case cohort ( n = 48 hospitalized patients from Heidelberg) as well as n = 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.
【저자키워드】 SARS-CoV-2, multiplex serology, 【초록키워드】 SARS-CoV-2 pandemic, Antibody Response, SARS-CoV-1, ELISA, Spike protein, Disease progression, Protein, sensitivity, specificity, Cohort, Antibody responses, Receptor-binding domain, Serological assay, pathogen, Immunoglobulin, Sensitivity and specificity, serological assays, Algorithm, Control, age, Pathogens, proteome, common cold, immunofluorescence, SARS-CoV-2 proteins, SARS-CoV-2 epidemiology, COVID-19 patient, dual, symptom onset, E. coli, COVID-19 case, SARS-CoV-2 proteome, SARS-CoV-2 research, fluorescent, SARS-CoV-2 protein, ccCoVs, common cold Coronaviruses, HEK293 cells, Ig isotypes, SARS-CoV-2 N, Research question, enzyme-linked immunosorbent, Course, assays, determine, hospitalized patient, the spike protein, expressed, analysis, the receptor-binding domain, Ig isotype, the SARS-CoV-2, 【제목키워드】 response, novel, the SARS-CoV-2,