Publication date: Jul 22, 2025

Choice of immunoassay influences population seroprevalence estimates. Post-hoc adjustments for assay performance could improve comparability of estimates across studies and enable pooled analyses. We assessed post-hoc adjustment methods using data from 2021-2023 SARS-CoV-2 serosurveillance studies in Alberta, Canada: one that tested 124,008 blood donations using Roche immunoassays (SARS-CoV-2 nucleocapsid total antibody and anti-SARS-CoV-2 S) and another that tested 214,780 patient samples using Abbott immunoassays (SARS-CoV-2 IgG and anti-SARS-CoV-2 S). Comparing datasets, seropositivity for antibodies against nucleocapsid (anti-N) diverged after May 2022 due to differential loss of sensitivity as a function of time since infection. The commonly used Rogen-Gladen adjustment did not reduce this divergence. Regression-based adjustments using the assays’ semi-quantitative results produced more similar estimates of anti-N seroprevalence and rolling incidence proportion (proportion of individuals infected in recent months). Seropositivity for antibodies targeting SARS-CoV-2 spike protein was similar without adjustment, and concordance was not improved when applying an alternative, functional threshold. These findings suggest that assay performance substantially impacted population inferences from SARS-CoV-2 serosurveillance studies in the Omicron period. Unlike methods that ignore time-varying assay sensitivity, regression-based methods using the semi-quantitative assay resulted in increased concordance in estimated anti-N seropositivity and rolling incidence between cohorts using different assays.

Semantics

Original Article