Performance evaluation of microfluidic microplate-based fluorescent ELISA for qualitative detection of SARS-CoV-2-specific IgG and IgM

Sci Rep. 2024 Aug 6;14(1):18200. doi: 10.1038/s41598-024-67977-9.

Abstract

We evaluated the diagnostic performance of newly developed microfluidic microplate-based fluorescent ELISA for anti-SARS-CoV-2 antibody detection: the Veri-Q opti COVID-19 IgG and IgM ELISAs (hereafter, "Opti IgG/M"; MiCo BioMed, Gyeonggi-do, Republic of Korea), in comparison with conventional ELISAs. A total of 270 serum samples were analyzed, among which 90 samples were serially obtained from 25 COVID-19 patients. Another 180 samples were collected from 180 SARS-CoV-2-negative individuals. As comparative assays, we used SCoV-2 Detect IgG/M ELISA (hereafter, "InBios IgG/M"; InBios, Seattle, WA, USA) and Veri-Q COVID-19 IgG/IgM ELISA (hereafter, "Veri-Q IgG/M"; MiCo BioMed). Compared with conventional ELISAs, the Opti IgG yielded 97.1-100.0% positive percent agreement, 95.2-98.0% negative percent agreement, 96.3-97.8% total percent agreement, and kappa values of 0.90-0.94. Between the Opti IgM and the InBios IgM, the values were 93.7%, 96.6%, 95.9%, and 0.89, respectively. For the Opti IgG, sensitivities for the samples collected from 0-7, 8-14, 15-21, and ≥ 22 days after symptom onset were 40.0, 58.3, 94.1, and 100.0%, respectively. The values for the Opti IgM were 30.0, 54.2, 88.2, and 80%, respectively. The diagnostic specificities of the Opti IgG and IgM were 99.4 and 97.2%, respectively. The microfluidic microplate-based fluorescent ELISAs showed comparable diagnostic performance to conventional ELISAs for detecting anti-SARS-CoV-2 antibodies. With the combination of high throughput, a simplified workflow, and the ability to analyze reduced volumes, this new technology has great potential for improving SARS-CoV-2 serologic testing.

Keywords: Antibody testing; COVID-19; ELISA; Microfluidic ELISA; SARS-CoV-2; Serologic assay.

Publication types

  • Evaluation Study

MeSH terms

  • Aged
  • Antibodies, Viral* / blood
  • Antibodies, Viral* / immunology
  • COVID-19 Serological Testing* / methods
  • COVID-19* / blood
  • COVID-19* / diagnosis
  • Enzyme-Linked Immunosorbent Assay* / methods
  • Female
  • Humans
  • Immunoglobulin G* / blood
  • Immunoglobulin M* / blood
  • Immunoglobulin M* / immunology
  • Male
  • Microfluidics / instrumentation
  • Microfluidics / methods
  • Middle Aged
  • SARS-CoV-2* / immunology
  • SARS-CoV-2* / isolation & purification
  • Sensitivity and Specificity

Substances

  • Immunoglobulin M
  • Immunoglobulin G
  • Antibodies, Viral