Evaluation of a novel multiplexed assay for determining IgG levels and functional activity to SARS-CoV-2

J Clin Virol. 2020 Sep:130:104572. doi: 10.1016/j.jcv.2020.104572. Epub 2020 Aug 2.

Abstract

Background: The emergence of SARS-CoV-2 has led to the development of serological assays that could aid in an understanding of the burden of COVID-19 disease. Many available tests lack rigorous evaluation and therefore results may be misleading.

Objectives: The aim of this study was to assess the performance of a novel multiplexed immunoassay for the simultaneous detection of antibodies against SARS-CoV-2 trimeric spike (S), spike receptor binding domain (RBD), spike N terminal domain and nucleocapsid antigen and a novel pseudo-neutralisation assay.

Methods: A multiplexed solid-phase chemiluminescence assay (Meso Scale Discovery) was evaluated for the simultaneous detection of IgG binding to four SARS-CoV-2 antigens and the quantification of antibody-induced ACE-2 binding inhibition (pseudo-neutralisation assay). Sensitivity was evaluated with a total of 196 COVID-19 serum samples (169 confirmed PCR positive and 27 anti-nucleocapsid IgG positive) from individuals with mild symptomatic or asymptomatic disease. Specificity was evaluated with 194 control serum samples collected from adults prior to December 2019.

Results: The specificity and sensitivity of the binding IgG assay was highest for S protein with a specificity of 97.4 % and sensitivity of 96.2 % for samples taken 14 days and 97.9 % for samples taken 21 days following the onset of symptoms. IgG concentration to S and RBD correlated strongly with percentage inhibition measured by the pseudo-neutralisation assay.

Conclusion: Excellent sensitivity for IgG detection was obtained over 14 days since onset of symptoms for three SARS-CoV-2 antigens (S, RBD and N) in this multiplexed assay which can also measure antibody functionality.

Keywords: Covid19; Immunoassays; Immunology; SARS-CoV-2.

Publication types

  • Evaluation Study

MeSH terms

  • Adult
  • Antibodies, Viral / blood*
  • Antigens, Viral / immunology*
  • Betacoronavirus
  • COVID-19
  • COVID-19 Testing
  • Clinical Laboratory Techniques
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / immunology
  • Coronavirus Nucleocapsid Proteins
  • Female
  • Humans
  • Immunoassay / methods*
  • Immunoglobulin G / blood*
  • Luminescent Measurements / methods
  • Male
  • Middle Aged
  • Nucleocapsid Proteins / immunology
  • Pandemics
  • Phosphoproteins
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / immunology
  • SARS-CoV-2
  • Sensitivity and Specificity
  • Spike Glycoprotein, Coronavirus / immunology

Substances

  • Antibodies, Viral
  • Antigens, Viral
  • Coronavirus Nucleocapsid Proteins
  • Immunoglobulin G
  • Nucleocapsid Proteins
  • Phosphoproteins
  • Spike Glycoprotein, Coronavirus
  • nucleocapsid phosphoprotein, SARS-CoV-2
  • spike protein, SARS-CoV-2