Evaluation of two commercial and two non-commercial immunoassays for the detection of prior infection to SARS-CoV-2

Nilles, Eric J.; Karlson, Elizabeth W.; Norman, Maia; Gilboa, Tal; Fischinger, Stephanie; Atyeo, Caroline; Zhou, Guohai; Bennett, Christopher L.; Tolan, Nicole V.; Oganezova, Karina; Walt, David R.; Alter, Galit; Simmons, Daimon P.; Schur, Peter; Jarolim, Petr; Baden, Lindsey R.

BACKGROUND: Seroepidemiology is an important tool to characterize the epidemiology and immunobiology of SARS-CoV-2 but many immunoassays have not been externally validated raising questions about reliability of study findings. To ensure meaningful data, particularly in a low seroprevalence population, assays need to be rigorously characterized with high specificity. METHODS: We evaluated two commercial (Roche Diagnostics and Epitope Diagnostics IgM/IgG) and two non-commercial (Simoa and Ragon/MGH IgG) immunoassays against 68 confirmed positive and 232 pre-pandemic negative controls. Sensitivity was stratified by time from symptom onset. The Simoa multiplex assay applied three pre-defined algorithm models to determine sample result. RESULTS: The Roche and Ragon/MGH IgG assays each registered 1/232 false positive, the primary Simoa model registered 2/232 false positives, and the Epitope registered 2/230 and 3/230 false positives for the IgG and IgM assays respectively. Sensitivity >21 days post symptom-onset was 100% for all assays except Epitope IgM, but lower and/or with greater variability between assays for samples collected 9–14 days (67–100%) and 15–21 days (69–100%) post-symptom onset. The Simoa and Epitope IgG assays demonstrated excellent sensitivity earlier in the disease course. The Roche and Ragon/MGH assays were less sensitive during early disease, particularly among immunosuppressed individuals. CONCLUSIONS: The Epitope IgG demonstrated good sensitivity and specificity. The Roche and Ragon/MGH IgG assays registered rare false positives with lower early sensitivity. The Simoa assay primary model had excellent sensitivity and few false positives.