Enzyme-linked immunosorbent assay (ELISA) for IgM, IgG and IgA class antibodies against<i>Candida albicans</i>antigens: Development and comparison with other methods

Kostiala, Anja A. I.; Kostiala, I.

doi:10.1080/00362178185380191

Cited by 40 publications

(17 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Indirect ELISA was performed as described previously (Kostiala & Kostiala, 1981). Antigen (pHMIN or ptHMIN) was added (1 ìg per well) to 96-well microtitre plates (Nunc-Immuno Starwell, MaxiSorp Surface) in 100 ìl carbonate buffer (63 mM; pH 9 .…”

Section: Methodsmentioning

confidence: 99%

ELISA for early diagnosis of histoplasmosis

Guimarães

Pizzini

Guedes

et al. 2004

Journal of Medical Microbiology

View full text Add to dashboard Cite

An ELISA was developed and evaluated as a method for detecting antibodies against glycosylated and deglycosylated histoplasmin (HMIN). Sera from patients with histoplasmosis, paracoccidioidomycosis, sporotrichosis, coccidioidomycosis, aspergillosis, cryptococcosis and healthy donors were tested by ELISA against purified, deglycosylated histoplasmin (ptHMIN) and compared with purified, native (i.e. glycosylated) histoplasmin (pHMIN). Although cross-reactivity was not abolished when ptHMIN was used in the test, it was reduced (pHMIN ELISA 93 % versus ptHMIN ELISA 96 %). However, there were statistically significant differences between the sensitivities of these two methods for the detection of antibodies (pHMIN ELISA 57 % versus ptHMIN ELISA 92 %; P , 0 . 001) and between the efficiency of the methods (pHMIN ELISA 83 % versus ptHMIN ELISA 95 %; P , 0 . 001). These parameters compare better than previously published data relating to the use of treated HMIN in diagnostic ELISAs. Some of the reactivities of serum samples were compared by immunoblotting using deglycosylated HMIN and by immunodiffusion using the crude antigen. The results demonstrated that cross-reactions with heterologous sera in both ELISAs could also be observed in immunoblotting and arose from shared protein epitopes. These data suggest that ELISA using deglycosylated HMIN is a very sensitive diagnostic method and, by using commercially available antigen, it can be easily standardized and performed faster than previous Western blot-based tests using the same antigen. It provides a useful adjunct to existing methods of diagnosis that could be applied even in situations where laboratory facilities were relatively limited. INTRODUCTIONHistoplasmosis is a systemic fungal disease caused by Histoplasma capsulatum. The significance of histoplasmosis results from its worldwide distribution (Rios-Fabra et al., 1994;Wheat, 2001), its ability to mimic other serious disease entities (Goodwin et al., 1980;McKinsey et al., 1997) and its propensity to cause serious disseminated infection in immunocompromised patients (Bradsher, 1996;Goodwin et al., 1980;Wheat, 1996;Wheat & Kauffman, 2003).Definitive diagnosis has typically relied either on direct visualization of the organism in tissue and/or the isolation of the causative organism, which is time-consuming and lacking in sensitivity (Bullock, 1995;Kwon-Chung & Bennet, 1992;Wheat, 2001). The detection of patients' antibody responses offers a more rapid alternative to microbiological means of diagnosis, and the detection of host anti-H. capsulatum antibodies by immunodiffusion (ID) and complement fixation (CF) tests is often used (Bullock, 1995;Kaufman et al., 1997). Both the yeast and mycelial phases of the fungus produce a number of exoantigens in culture, the most important and characteristic being the H and M antigens. These two antigens are the primary immunoreactive constituents of histoplasmin (HMIN), the standard diagnostic reagent used in ID and CF for many years (Standard & Kaufman, 1976). However, it was...

show abstract

Section: Methodsmentioning

confidence: 99%

ELISA for early diagnosis of histoplasmosis

Guimarães

Pizzini

Guedes

et al. 2004

Journal of Medical Microbiology

View full text Add to dashboard Cite

show abstract

“…Serial twofold dilutions of sera in saline against a Candida albicans extract (1 in 10, w/v, lot G13821M, Hollister-Stier, Berkeley, Ca, USA) were studied by ID as described earlier (Kostiala and Kostiala, 1981). Titres of 3 1 were considered positive.…”

Section: Patientsmentioning

confidence: 99%

“…albicans (Pasteur Institute, Paris, France) were used in CIE as previously reported (Kostiala and Kostiala, 1981) to determine titres in serial twofold dilutions of sera made in saline. As a control, a positive reference serum (Anti-Candida albicans Serum, Pasteur Institute) with a titre of 32 against S and 4 against M antigens was used.…”

Section: Patientsmentioning

confidence: 99%

See 1 more Smart Citation

Antibodies against antigens of Candida albicans in patients with fungaemia and bacteraemia, studied by ELISA, precipitation, passive haemagglutination and immunofluorescence techniques

Kostiala

Larinkari

et al. 1981

Journal of Medical Microbiology

Self Cite

View full text Add to dashboard Cite

SUMMARY. Antibodies against commercially available antigens of Candida albicans were assayed in 54 sera from 24 patients with fungaemia and in 66 sera from 33 patients with bacteraemia. In patients with persistent fungaemia, antibody was found during the week after the fungus was first cultured from the blood, but peak titres did not usually occur until the end of the second week. A significant rise of titre in C. albicans infection was observed in 50% of paired sera tested by passive haemagglutination (PHA), indirect immunofluorescence (IF) and Ouchterlony immunodiffusion (ID). The same percentage was obtained by counterimmunoelectrophoresis (CIE) against candida metabolic antigens, whereas it was increased to 88% when somatic antigens were used. Enzymelinked immunosorbent assay (ELISA) demonstrated a rise of titre in 25,75 and 50% of sera in IgM, IgG and IgA assays, respectively. Sera from patients with transient fungaemia demonstrated persistent antibody titres.In paired sera from patients with bacteraemia, ID and CIE titres were low (G4). There was an increase of candida antibodies in 0-9% of patients by ELISA, ID or CIE and in 18-21% by PHA or IF. Clinically significant fungaemia was most reliably differentiated serologically from bacteraemia by CIE S-antigen and ELISA IgG assays.

show abstract

“…ELISA has been employed in the serology of various mycotic diseases (7,10,11,12,13,21,22). It is an extremely sensitive method and is reagent saving compared with other assays.…”

mentioning

confidence: 99%

Improvement of the Specificity of an Enzyme-Linked Immunosorbent Assay for Diagnosis of Paracoccidioidomycosis

2005

View full text Add to dashboard Cite

In an attempt to improve the specificity of an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of paracoccidioidomycosis (PCM), sera from patients with PCM were tested using various approaches, such as sodium metaperiodate antigen (gp43) treatment, a serum absorption process with Candida albicans or Histoplasma capsulatum antigens, and dilution of serum in galactose, the main common epitope among pathogenic fungi. The maximum specificity found in this ELISA was 84%. All of these procedures proved inefficient for eliminating all cross-reacting antibodies and obtaining an ELISA specific for PCM diagnosis.Paracoccidioidomycosis (PCM) is a mycotic disease caused by the dimorphic fungus Paracoccidioides brasiliensis and causes a deep mycosis, resulting in a severe chronic granulomatous infection of the skin, mucous membranes, lymph nodes, and internal organs. Almost all South and Central America countries have large regions where PCM is endemic. The definitive diagnosis of PCM includes direct observation of the characteristic multiple-budding cells in biological fluids and tissue sections or isolation of the fungus from clinical materials. For cases in which P. brasiliensis is not observed through direct examination, several serological tests have been used to detect antibodies against the fungus in order to establish the diagnosis (2). gp43, a 43,000-Da glycoprotein from P. brasiliensis, is a major diagnostic antigen in serological assays (1,3,15,18,23,24). It has been demonstrated that gp43 is as good as crude exoantigen preparations (concentrated, dialyzed culture supernatants) (3, 4) for detecting precipitating antibody in immunodiffusion tests, and purified gp43 has also been adapted to several other serological tests, such as immunoprecipitation of 125 I-labeled gp43 (IPP), conventional enzymelinked immunosorbent assay (ELISA), capture enzyme immunoassay (EIA), and passive hemagglutination (5,20,23). Cross reactivity in serological assays from patients with histoplasmosis depends on the test used and seems to involve a carbohydrate epitope. While IPP and capture EIA using gp43 appear specific for PCM diagnosis, experiments were performed to determine the nature of the cross reactivity in sera from those patients with histoplasmosis that reacted with gp43 in the IPP tests. Treatment of gp43 with 0.2 M D-galactose inhibited such cross reactivity, and no cross reactivity was observed with the deglycosylated form of gp43, with EH38, or with the 38,000-Da protein which had been treated with periodate (18). Owing to the N-linked high-mannose nature of the gp43 glycan, deglycosylation was efficiently carried out with endo-␤-N-acetylglucosaminidase H. Since sera from patients with histoplasmosis generally reacted with the high-molecular-weight glycocomplex present in the crude antigen of P. brasiliensis, this reactivity and that with gp43 likely involved recognition of terminal ␤-galactofuranosyl units (24).Travassos et al. (24) related that in general, specific reactions with gp43 are obtained when the ant...

show abstract

Enzyme-linked immunosorbent assay (ELISA) for IgM, IgG and IgA class antibodies againstCandida albicansantigens: Development and comparison with other methods

Cited by 40 publications

References 14 publications

ELISA for early diagnosis of histoplasmosis

ELISA for early diagnosis of histoplasmosis

Antibodies against antigens of Candida albicans in patients with fungaemia and bacteraemia, studied by ELISA, precipitation, passive haemagglutination and immunofluorescence techniques

Improvement of the Specificity of an Enzyme-Linked Immunosorbent Assay for Diagnosis of Paracoccidioidomycosis

Contact Info

Product

Resources

About