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Diagnostic value of anti-Candida enolase antibodies
A J van Deventer1, H J van Vliet, W C Hop
1Department of Clinical Microbiology and Antimicrobial Therapy, Erasmus University Rottedam, The Netherlands.
Insights
A new assay using Candida enolase antigen shows promise in distinguishing invasive candidiasis from colonization. This diagnostic tool offers significant discrimination in both immunocompetent and immunodeficient patients.
Area of Science:
- Medical Microbiology
- Immunology
- Diagnostic Development
Background:
- Invasive candidiasis is a serious infection, and differentiating it from colonization is crucial for effective treatment.
- Current diagnostic methods can be limited in accuracy and speed.
- Candida enolase is an immunodominant antigen identified in Candida species.
Purpose of the Study:
- To develop and evaluate an assay for detecting antibodies against Candida enolase to differentiate invasive candidiasis from colonization.
- To compare the diagnostic performance of an enzyme-linked immunosorbent assay (ELISA) with counterimmunoelectrophoresis (CIE).
Main Methods:
- Purification of an immunodominant antigen with enolase enzyme activity from Candida.
- Development of an antigen (Au) enzyme-linked immunosorbent assay (ELISA) using the purified Candida enolase.
- Testing sera from patients with invasive candidiasis or Candida colonization using ELISA and CIE.
- Validation using Western blots and tandem crossed immunoelectrophoresis.
Main Results:
- The Au ELISA significantly discriminated between invasive candidiasis and colonization in both immunocompetent (50% sensitivity, 86% specificity) and immunodeficient (53% sensitivity, 78% specificity) patients.
- The CIE assay showed higher sensitivity but lower specificity in immunocompetent patients and failed to discriminate in immunodeficient patients.
- Increasing serum volume did not significantly improve assay sensitivity or specificity.
- Combining antibody assays with antigen detection (Cand-Tec) did not enhance predictive value for invasive candidiasis.
Conclusions:
- The Candida enolase antigen-based ELISA is a valuable tool for distinguishing invasive candidiasis from colonization.
- The ELISA demonstrates a significant ability to differentiate infection states across different patient immune statuses.
- Further optimization may be needed, but the assay shows potential for improved Candida diagnostics.
Abstract:
An immunodominant antigen with enolase enzyme activity was purified and used for the development of an assay to detect antibodies directed against this antigen in sera from patients with either invasive candidiasis or Candida colonization. The Au enzyme-linked immunosorbent assay established with the Candida enolase antigen was able to discriminate significantly between invasive candidiasis and colonization in both immunocompetent and immunodeficient groups of patients. The test had a sensitivity of 50% and a specificity of 86% in the immunocompetent patient group. In the immunodeficient patient group, a sensitivity of 53% and a specificity of 78% were established. Antibody levels determined by a counterimmunoelectrophoresis assay with the same set of sera resulted in a better sensitivity for sera from the immunocompetent patient group but a lower specificity, i.e., 80 and 29%, respectively. The counterimmunoelectrophoresis assay of sera from the immunodeficient patient group was not able to discriminate significantly between invasive candidiasis and colonization. With the use of more serum from each patient, the sensitivity of the antibody detection assays increased, while the specificity was maintained. The increase, however, was not statistically significant. Combining the results of the antibody assays with antigen titers obtained by the Cand-Tec assay did not improve the predictive value with respect to invasive candidiasis, as determined by multivariance regression analysis. Furthermore, it was demonstrated by performance of Western blots (immunoblots) that sera from patients as well as a rabbit antiserum cross-reacted with the Candida enolase and baker's yeast enolase enzyme. However, by tandem crossed immunoelectrophoresis it was demonstrated that the antibodies were directed toward different epitopes of the antigen.
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