Microscopy-based assay for semi-quantitative detection of SARS-CoV-2 specific antibodies in human sera: A semi-quantitative, high throughput, microscopy-based assay expands existing approaches to measure SARS-CoV-2 specific antibody levels in human sera

Bioessays. 2021 Mar;43(3):e2000257. doi: 10.1002/bies.202000257. Epub 2020 Dec 30.

Abstract

Emergence of the novel pathogenic coronavirus SARS-CoV-2 and its rapid pandemic spread presents challenges that demand immediate attention. Here, we describe the development of a semi-quantitative high-content microscopy-based assay for detection of three major classes (IgG, IgA, and IgM) of SARS-CoV-2 specific antibodies in human samples. The possibility to detect antibodies against the entire viral proteome together with a robust semi-automated image analysis workflow resulted in specific, sensitive and unbiased assay that complements the portfolio of SARS-CoV-2 serological assays. Sensitive, specific and quantitative serological assays are urgently needed for a better understanding of humoral immune response against the virus as a basis for developing public health strategies to control viral spread. The procedure described here has been used for clinical studies and provides a general framework for the application of quantitative high-throughput microscopy to rapidly develop serological assays for emerging virus infections.

Keywords: SARS-CoV-2; antibody; immunofluorescence; machine learning image analysis; quantitative microscopy; serological test.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Viral / blood*
  • COVID-19 / diagnosis*
  • COVID-19 / immunology
  • COVID-19 / virology
  • COVID-19 Testing / methods
  • Fluorescent Antibody Technique
  • High-Throughput Screening Assays
  • Humans
  • Image Processing, Computer-Assisted / statistics & numerical data
  • Immune Sera / chemistry
  • Immunoassay*
  • Immunoglobulin A / blood*
  • Immunoglobulin G / blood*
  • Immunoglobulin M / blood*
  • Machine Learning
  • Microscopy / methods*
  • SARS-CoV-2 / immunology*
  • Sensitivity and Specificity

Substances

  • Antibodies, Viral
  • Immune Sera
  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin M