Systematic evaluation of SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay.
JSON
Hober S, Hellström C, Olofsson J, Andersson E, Bergström S, Jernbom Falk A, Bayati S, Mravinacova S, Sjöberg R, Yousef J, Skoglund L, Kanje S, Berling A, Svensson AS, Jensen G, Enstedt H, Afshari D, Xu LL, Zwahlen M, von Feilitzen K, Hanke L, Murrell B, McInerney G, Karlsson Hedestam GB, Lendel C, Roth RG, Skoog I, Svenungsson E, Olsson T, Fogdell-Hahn A, Lindroth Y, Lundgren M, Maleki KT, Lagerqvist N, Klingström J, Da Silva Rodrigues R, Muschiol S, Bogdanovic G, Arroyo Mühr LS, Eklund C, Lagheden C, Dillner J, Sivertsson Å, Havervall S, Thålin C, Tegel H, Pin E, Månberg A, Hedhammar M, Nilsson P
Clin Transl Immunol 10 (7) e1312
[2021-07-19; online 2021-07-19]
The COVID-19 pandemic poses an immense need for accurate, sensitive and high-throughput clinical tests, and serological assays are needed for both overarching epidemiological studies and evaluating vaccines. Here, we present the development and validation of a high-throughput multiplex bead-based serological assay. More than 100 representations of SARS-CoV-2 proteins were included for initial evaluation, including antigens produced in bacterial and mammalian hosts as well as synthetic peptides. The five best-performing antigens, three representing the spike glycoprotein and two representing the nucleocapsid protein, were further evaluated for detection of IgG antibodies in samples from 331 COVID-19 patients and convalescents, and in 2090 negative controls sampled before 2020. Three antigens were finally selected, represented by a soluble trimeric form and the S1-domain of the spike glycoprotein as well as by the C-terminal domain of the nucleocapsid. The sensitivity for these three antigens individually was found to be 99.7%, 99.1% and 99.7%, and the specificity was found to be 98.1%, 98.7% and 95.7%. The best assay performance was although achieved when utilising two antigens in combination, enabling a sensitivity of up to 99.7% combined with a specificity of 100%. Requiring any two of the three antigens resulted in a sensitivity of 99.7% and a specificity of 99.4%. These observations demonstrate that a serological test based on a combination of several SARS-CoV-2 antigens enables a highly specific and sensitive multiplex serological COVID-19 assay.
Funder: KAW/SciLifeLab National COVID program
Research Area: Biobanks for COVID-19 research
Research Area: High-throughput and high-content serology
Research Area: Host cell systems biology and targets
PubMed 34295471
DOI 10.1002/cti2.1312
Crossref 10.1002/cti2.1312
pmc: PMC8288725
pii: CTI21312