SARS-CoV-2 Serologic Assays in Control and Unknown Populations Demonstrate the Necessity of Virus Neutralization Testing

J Infect Dis. 2021 Apr 8;223(7):1120-1131. doi: 10.1093/infdis/jiaa797.

Abstract

Background: To determine how serologic antibody testing outcome links with virus neutralization of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), we evaluated individuals for SARS-CoV-2 antibody level and viral neutralization.

Methods: We compared serum Ig levels across platforms of viral antigens and antibodies with 15 positive and 30 negative SARS-CoV-2 controls followed by viral neutralization assessment. We then applied these platforms to a clinically relevant cohort of 114 individuals with unknown histories of SARS-CoV-2 infection.

Results: In controls, the best-performing virus-specific antibody detection platforms were SARS-CoV-2 receptor binding domain (RBD) IgG (sensitivity 87%, specificity 100%, positive predictive value [PPV] 100%, negative predictive value [NPV] 94%), spike IgG3 (sensitivity 93%, specificity 97%, PPV 93%, NPV 97%), and nucleocapsid protein (NP) IgG (sensitivity 93%, specificity 97%, PPV 93%, NPV 97%). Neutralization of positive and negative control sera showed 100% agreement. Twenty individuals with unknown history had detectable SARS-CoV-2 antibodies with 16 demonstrating virus neutralization. Spike IgG3 provided the highest accuracy for predicting serologically positive individuals with virus neutralization activity (misidentified 1/20 unknowns compared to 2/20 for RBD and NP IgG).

Conclusions: The coupling of virus neutralization analysis to a spike IgG3 antibody test is optimal to categorize patients for correlates of SARS-CoV-2 immune protection status.

Keywords: SARS-CoV-2; antibody; immunity; neutralization; serologic; two-tiered testing.

Publication types

  • Comparative Study
  • Observational Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies, Neutralizing / blood
  • Antibodies, Neutralizing / immunology
  • Antibodies, Neutralizing / isolation & purification
  • Antibodies, Viral / blood
  • Antibodies, Viral / immunology
  • Antibodies, Viral / isolation & purification
  • COVID-19 / blood
  • COVID-19 / diagnosis*
  • COVID-19 / epidemiology
  • COVID-19 / immunology
  • COVID-19 Serological Testing / methods*
  • Enzyme-Linked Immunosorbent Assay
  • False Positive Reactions
  • Female
  • Humans
  • Immunity, Humoral
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Immunoglobulin G / isolation & purification
  • Male
  • Middle Aged
  • Neutralization Tests / methods*
  • Predictive Value of Tests
  • Prevalence
  • SARS-CoV-2 / immunology*
  • SARS-CoV-2 / isolation & purification
  • Sensitivity and Specificity
  • Seroepidemiologic Studies
  • Spike Glycoprotein, Coronavirus / immunology
  • Young Adult

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Immunoglobulin G
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2