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Simultaneous Measurement of IgM and IgG Antibodies to SARS-CoV-2 Spike, RBD, and Nucleocapsid Multiplexed in a Single Assay on the xMAP INTELLIFLEX DR-SE Flow Analyzer.
Cameron, Andrew; Bohrhunter, Jessica L; Porterfield, Claire A; Mangat, Rupinder; Karasick, Michael H; Pearson, Zachary; Angeloni, Stephen; Pecora, Nicole D.
  • Cameron A; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Bohrhunter JL; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Porterfield CA; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Mangat R; Division of Infectious Diseases, Department of Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Karasick MH; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Pearson Z; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
  • Angeloni S; Luminex Corporation, Austin, Texas, USA.
  • Pecora ND; Pathology and Laboratory Medicine, University of Rochester Medical Centergrid.412750.5, Rochester, New York, USA.
Microbiol Spectr ; 10(2): e0250721, 2022 04 27.
Article in English | MEDLINE | ID: covidwho-1779319
ABSTRACT
The multiplex capabilities of the new xMAP INTELLIFLEX DR-SE flow analyzer were explored by modifying a serological assay previously used to characterize the IgG antibody to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The goal was to examine the instrument's performance and to simultaneously measure IgM and IgG antibody responses against multiple SARS-CoV-2 antigens in a single assay. Specific antibodies against the SARS-CoV-2 spike (S), receptor binding domain (RBD), and nucleocapsid (N) proteins were investigated in 310 symptomatic case patients using a fluorescent microsphere immunoassay and simultaneous detection of IgM and IgG. Neutralization potential was studied using the addition of soluble angiotensin-converting enzyme 2 (ACE2) to block antibody binding. A profile extending to 180 days from symptom onset (DFSO) was described for antibodies specific to each viral antigen. Generally, IgM levels peaked and declined rapidly ∼3-4 weeks following infection, whereas S- and RBD-specific IgG plateaued at 80 DFSO. ACE2 more effectively prevented IgM and IgG binding in convalescent cases > 30 DFSO, suggesting those antibodies had greater neutralization potential. This work highlighted the multiplex and multi-analyte potential of the xMAP INTELLIFLEX DR-SE, and provided further evidence for antigen-specific IgM and IgG trajectories in acute and convalescent cases. IMPORTANCE The xMAP INTELLIFLEX DR-SE enabled simultaneous and semi-quantitative detection of both IgM and IgG to three different SARS-CoV-2 antigens in a single assay. The assay format is advantageous for rapid and medium-throughput profiling using a small volume of specimen. The xMAP INTELLIFLEX DR-SE technology demonstrated the potential to include numerous SARS-CoV-2 antigens; future work could incorporate multiple spike protein variants in a single assay. This could be an important feature for assessing the serological response to emerging variants of SARS-CoV-2.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Topics: Variants Limits: Humans Language: English Journal: Microbiol Spectr Year: 2022 Document Type: Article Affiliation country: Spectrum.02507-21

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Topics: Variants Limits: Humans Language: English Journal: Microbiol Spectr Year: 2022 Document Type: Article Affiliation country: Spectrum.02507-21