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Evaluation of an enzyme immunoassay using neoglycolipids constructed fromCandida albicans oligomannosides to define the specificity of anti-mannan antibodies

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Abstract

In order to study the respective roles of oligomannoside sequences in the antigenicity ofCandida albicans phosphopeptidomannan, a method was developed for constructing neoglycolipids from oligomannosides released by depolymerisation of this molecule. Oligomannosides released by acetolysis were converted to neoglycolipids by coupling them to 4-hexadecylaniline in an equimolar reaction checked by thin layer chromatography. When coated onto microEIA plates, the neoglycolipids exhibited strong reactions which were dose dependent and were saturable with concanavalin A. Reactivity of neoglycolipids with immunoglobulins were then tested with a panel of monoclonal and polyclonal antibodies reacting with epitopes present in the original phosphopeptidomannan. One of two IgM monoclonal antibodies and two of five monospecific rabbit polyclonal IgG reacted strongly with neoglycolipids therefore providing evidence of the presence of structures mimicking epitopes within the pool of neoglycolipids. When 38 sera from 18 hospital inpatients with various levels of antibodies toCandida albicans were tested, a correlation was observed between the EIA to detect neoglycolipids and the EIA to detect phosphopeptidomannan. Successive sera from all patients showing seroconversion in the immunofluorescence assay had increased EIA signals for neoglycolipids.

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Authors and Affiliations

  1. Institut National de la Santé et de la Recherche Médicale, U 42, Domaine du CERTIA, 369, Rue J. Guesde, BP 39, 59650, Villeneuve d'Ascq, France

    C. Faille & D. Poulain

  2. Mycological Reference Laboratory, Central Public Health Laboratory Service, 61 Colindale Avenue, NW95HT, London, UK

    D. W. R. Mackenzie

  3. Laboratoire de Chimie Biologique et Unité Mixte Centre national de la Recherche Scientifique 111, Université des Sciences et Techniques de Lille, Flandres-Artois, 59650, Villeneuve d'Ascq Cedex, France

    J. C. Michalski

  4. Service de Parasitologie-Mycologie, Faculté de Médecine, 59000, Lille, France

    D. Poulain

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  1. C. Faille
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  2. D. W. R. Mackenzie
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  3. J. C. Michalski
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  4. D. Poulain
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Faille, C., Mackenzie, D.W.R., Michalski, J.C. et al. Evaluation of an enzyme immunoassay using neoglycolipids constructed fromCandida albicans oligomannosides to define the specificity of anti-mannan antibodies. Eur. J. Clin. Microbiol. Infect. Dis. 11, 438–446 (1992). https://doi.org/10.1007/BF01961859

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