A quantitative enzyme-linked immunoassay (ELISA) to approximate complement-fixing antibody titers in serum from patients with coccidioidomycosis

Tao Peng, Yue Zong, Michael DL Johnson, Sanjay V. Menghani, Maria Lourdes Lewis, John N. Galgiani

Research output: Contribution to journalArticlepeer-review

Abstract

Coccidioidomycosis is most frequently diagnosed serologically, and the quantitative test for complement-fixing antibodies is considered prognostically useful. Because complement-fixing antibody testing is complex, labor-intensive, and poorly standardized, an enzyme-linked immunoassay (ELISA) alternative would be attractive. In this report, we restrict the complement-fixing, antibody-binding domain to a 200-amino-acid recombinant peptide of the known antigen. Over-lapping truncations of this peptide do not bind complement-fixing antibodies, suggesting that the responsible epitope(s) are conformational. Further, anchoring the antigenic peptide to the ELISA plate by means of a C-terminal biotin-mimic peptide tag instead of allowing the peptide to randomly adhere to the plastic plate improves sensitivity of antibody detection by 1–2 logs in different sera. The newly developed ELISA shows a significant quantitative correlation with complement-fixing antibody titers. This ELISA shows potential as the basis for a new quantitative assay for coccidioidal antibodies.

Original languageEnglish (US)
Article number115198
JournalDiagnostic Microbiology and Infectious Disease
Volume99
Issue number1
DOIs
StatePublished - Jan 2021

Keywords

  • Antibodies
  • Coccidioidomycosis
  • Diagnostic test
  • ELISA
  • Epitopes
  • Recombinant proteins

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

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