title: Diagnosis of systemic candidiasis by enzyme immunoassay detection of specific antibodies to mycelial phase cell wall and cytoplasmic candidal antigens.
abstract: Diagnosis of systemic Candida infections was attempted by the use of an enzyme-linked immunosorbent assay (EIA) to detect IgG antibodies towards cell wall-bound and cytoplasmic candidal antigens. Cell wall antigens were sequentially solubilized by treatment of germinated blastoconidia of Candida albicans (ATCC 26555 strain) with beta-mercaptoethanol (beta ME extract) and digestion with Zymolyase 20T, a beta-glucanase preparation (Zymolyase extract). Protoplasts obtained after treatment with Zymolyase were osmotically lysed (cytoplasmic antigens). Sera were obtained from patients with systemic (n = 28) and superficial (n = 46) candidiasis. Control sera were obtained from normal healthy individuals (n = 31) and from hospitalized patients at low (n = 36) and at high (n = 13) risk of developing systemic candidiasis yet showing no symptoms of candidal infection. Detection of antibodies in crude sera samples by EIA using all of these antigenic extracts was highly specific (98-100%), but sensitivity of the method was low (3.5-17.8%). However, adsorption of sera with latex microspheres coated with purified Candida mannan in order to selectively remove anti-mannan antibodies prior to EIA improved the diagnostic efficiency of this test. Improvement was particularly noticeable when the beta ME extract was used as antigenic preparation, yielding a sensitivity of 89.2% and a specificity of 98.6%.
authors.affiliation: Unidad Departamental de Microbiología, Facultad de Medicina, Universitat de Valencia, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Navarro
authors.forename: E
authors.initials: E
authors.lastname: Monzonis
authors.forename: J L
authors.initials: JL
authors.lastname: López-Ribot
authors.forename: P
authors.initials: P
authors.lastname: Sepúlveda
authors.forename: M
authors.initials: M
authors.lastname: Casanova
authors.forename: J M
authors.initials: JM
authors.lastname: Nogueira
authors.forename: J P
authors.initials: JP
authors.lastname: Martínez
mesh_terms: D000908:Antibodies, Fungal / Q000097:blood*; D000946:Antigens, Fungal / Q000737:chemistry*; D015153:Blotting, Western; D002176:Candida albicans / Q000254:growth & development / Q000276:immunology* / Q000302:isolation & purification; D002177:Candidiasis / Q000175:diagnosis* / Q000276:immunology; D002473:Cell Wall / Q000276:immunology; D003593:Cytoplasm / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000032:analysis / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007841:Latex Fixation Tests; D008351:Mannans / Q000276:immunology; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D005656:Fungal Proteins; D007074:Immunoglobulin G; D008351:Mannans; D008562:Membrane Glycoproteins; C025363:mannoproteins