Stringent thresholds in SARS-CoV-2 IgG assays lead to under-detection of mild infections

BMC Infect Dis. 2021 Feb 18;21(1):187. doi: 10.1186/s12879-021-05878-2.

Abstract

Background: Thresholds for SARS-CoV-2 antibody assays have typically been determined using samples from symptomatic, often hospitalised, patients. In this setting the sensitivity and specificity of the best performing assays can both exceed 98%. However, antibody assay performance following mild infection is less clear.

Methods: We assessed quantitative IgG responses in a cohort of healthcare workers in Oxford, UK, with a high pre-test probability of Covid-19, in particular the 991/11,475(8.6%) who reported loss of smell/taste. We use anosmia/ageusia and other risk factors as probes for Covid-19 infection potentially undiagnosed by immunoassays by investigating their relationship with antibody readings either side of assay thresholds.

Results: The proportion of healthcare workers reporting anosmia/ageusia increased at antibody readings below diagnostic thresholds using an in-house ELISA (n = 9324) and the Abbott Architect chemiluminescent microparticle immunoassay (CMIA; n = 11,324): 426/906 (47%) reported anosmia/ageusia with a positive ELISA, 59/449 (13.1%) with high-negative and 326/7969 (4.1%) with low-negative readings. Similarly, by CMIA, 518/1093 (47.4%) with a positive result reported anosmia/ageusia, 106/686 (15.5%) with a high-negative and 358/9563 (3.7%) with a low-negative result. Adjusting for the proportion of staff reporting anosmia/ageusia suggests the sensitivity of both assays in mild infection is lower than previously reported: Oxford ELISA 89.8% (95%CI 86.6-92.8%) and Abbott CMIA 79.3% (75.9-82.7%).

Conclusion: Following mild SARS-CoV-2 infection 10-30% of individuals may have negative immunoassay results. While lowered diagnostic thresholds may result in unacceptable specificity, our findings have implications for epidemiological analyses and result interpretation in individuals with a high pre-test probability. Samples from mild PCR-confirmed infections should be included in SARS-CoV-2 immunoassay evaluations.

Keywords: Ageusia; Anosmia; Antibodies; COVID-19; SARS-CoV-2; Serology.

MeSH terms

  • Adult
  • Ageusia / virology
  • Anosmia / virology
  • Antibodies, Viral / analysis*
  • Asymptomatic Infections
  • COVID-19 / diagnosis*
  • COVID-19 Serological Testing / standards*
  • Enzyme-Linked Immunosorbent Assay / standards
  • Female
  • Health Personnel
  • Humans
  • Immunoassay / standards
  • Immunoglobulin G / analysis*
  • Male
  • Middle Aged
  • Sensitivity and Specificity
  • Undiagnosed Diseases
  • United Kingdom

Substances

  • Antibodies, Viral
  • Immunoglobulin G