Comparison of antibody measurements against Aspergillus fumigatus by means of double-diffusion and enzyme-linked immunosorbent assay (ELISA)

Kauffman, Henk F.; Beaumont, F; Heide, S. van der; Vries, K. de

doi:10.1016/0091-6749(83)90029-5

Cited by 69 publications

(51 citation statements)

References 13 publications

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“…The most commonly used methods are agar gel double immunodiffusion [16-18, 22, 29], crossed immunoelectrophoresis [17], ELISA [8,9,17,29] and CRIE [17,34]. However, the results obtained by these methods have been inconsistent due to differences in the sensitivity of the detection methods and the lack of standardization of antigen preparations.…”

Section: Discussionmentioning

confidence: 99%

Localization, Molecular Weight and Immunoglobulin Subclass Response to <i>Aspergillus fumigatus</i> Allergens in Acute Bronchopulmonary Aspergillosis

Leung

Gershwin

Coppel

et al. 1988

Int Arch Allergy Immunol

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A specimen of Aspergillus fumigatus was isolated from a patient with acute bronchopulmonary aspergillosis (ABPA). Cultures were allowed to propagate and were separated into spores and mycelium to > 95% homogeneity. Extracts of both the spores and mycelium were prepared and used for study by both dot blot analysis and immunoblotting to study the major allergens. By dot blot analysis, 22 of 22 patients with ABPA and none of 10 healthy controls reacted with mycelium when probed for both IgG and IgE anti-Aspergillus reactivity. Similar results were obtained when these extracts were separated on polyacrylamide gel electrophoresis and then probed; this included both IgG and IgE reactivity. Further, the reactivity to preparations of whole extract was absorbed with mycelium but not spores. Several distinct protein bands were detected with IgG, ranging from 30 to 110 kilodaltons (kD). In contrast, the majority of patients with ABPA, when studied for IgE reactivity, reacted only with a 70-kD protein, although 1 out of 22 patients reacted with a 30-kD protein. Subclass analysis of IgG reactivity demonstrated that 90% of reactive sera contained IgG2 and 73% IgG4 reactivity. IgG1 and IgG3 reactivity were present but less frequently detected. The use of immunoblotting will enable the identification of relevant allergens. Further, the demonstration that the allergenicity is located primarily in mycelium will allow more definitive studies of allergen isolation for studies of reactivity and, ultimately, cloning of the 70-kD protein and, finally, epitope mapping.

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Section: Discussionmentioning

confidence: 99%

Localization, Molecular Weight and Immunoglobulin Subclass Response to <i>Aspergillus fumigatus</i> Allergens in Acute Bronchopulmonary Aspergillosis

Leung

Gershwin

Coppel

et al. 1988

Int Arch Allergy Immunol

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“…Enzyme-linked immunosorbent assay Indirect ELISA was performed for the detection of total immunoglobulins against Aspergillus crude antigens according to the method described by Kauffman et al 23 with minor modifications as adopted in our previous studies.…”

Section: 3mentioning

confidence: 99%

Bronchogenic carcinoma and secondary aspergillosis—Common yet unexplored

Shahid

Malik

Bhargava

2008

Cancer

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BACKGROUND.More cases of bronchogenic carcinoma have been reported in recent years, and these patients are more prone to secondary aspergillosis. However, the frequency of secondary aspergillosis in bronchogenic carcinoma still has not been defined clearly in the literature.METHODS.The current study population was comprised of 69 patients with bronchogenic carcinoma and 16 healthy controls. Histopathologic examination was done to identify carcinoma cell types and to categorize aspergillosis types. Bronchoalveolar lavage (BAL) fluids were collected for direct fungal examination, culture, Aspergillus polymerase chain reaction (PCR), and galactomannan (GM) detection using a 1‐stage immunoenzymatic sandwich microplate assay; and blood samples were collected for fungal serology by double immunodiffusion (DID), enzyme‐linked immunosorbent assay (ELISA), and dot blot assay (DBA). The sensitivity, specificity, positive predictive value, and negative predictive value were analyzed for various nonvalidated tests. Twenty‐five patients had follow‐up data available for an analysis of clinical and diagnostic outcomes.RESULTS.The cohort included patients with squamous cell carcinoma (n = 47), adenocarcinoma (n = 16), small cell carcinoma (n = 3), large cell carcinoma (n = 2), and undiagnosed type (n = 1), and patients were categorized with definite invasive pulmonary aspergillosis (IPA) (n = 6), probable IPA (n = 17), possible IPA (n = 13), and non‐IPA (n = 33). Most patients were in the group ages 45 years to <60 years, and there was a preponderance of men (10.5:1). Cultures from 20 of 69 patients (29%) revealed the growth of Aspergillus species. Anti‐Aspergillus antibodies were detected in 26 of 69 patients (37.7%) each by DID and DBA, whereas antibodies were detected in 28 of 69 patients (40.6%) by ELISA. GM was detected in BAL fluids from 25 of 69 patients (36.2%), whereas Aspergillus DNA was detected in 32 of 69 patients (46.4%) by PCR. The sensitivity of PCR and serologic tests (ELISA, DID, and DBA) was 100% for definite IPA, whereas the sensitivity of PCR was comparatively higher than that of serologic tests for probable IPA, possible IPA, and non‐IPA.CONCLUSIONS.The current study indicated that there frequently is an association between bronchogenic carcinoma and secondary aspergillosis, and definite IPA and probable IPA are common clinical problems in patients with nonsmall cell lung cancer. Cancer 2008. © 2008 American Cancer Society.

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“…Existe evidencia que no todos los componentes antigénicos precipitantes se unen a la superficie del poliestireno y visceversa (27). Sin embargo, en este estudio no observamos diferencias significativas entre los resultados obtenidos en la DD, CIE+DD y ELISA.…”

Section: Materiales Y Métodosunclassified

“…La mayoría de los diagnósticos y estudios inmunológicos de aspergilosis por A. fumigatus, se centran principalmente en el uso de antígenos que son parte integral del micelio del hongo o de los productos extracelulares que demuestran propiedades de unión a IgG e IgE (16)(17)(18)(19) y aunque se han observado variabilidades aún entre lote y lote de un antígeno proveniente de una misma colonia (20)(21)(22)(23), esta variabilidad es menor en los antígenos que provienen del micelio roto que en los tradicionalmente preparados a partir de filtrado de cultivo, lo que se refleja en los resultados de las pruebas donde se utilizan dichos antígenos (24)(25)(26)(27).…”

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Extracción y caracterización de antígeno micelial de Aspergillus fumigatus

Cuadrado¹,

Gómez²

1995

biomedica

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ResumenEste estudio presenta la estandarización y caracterización antigénica de un extracto micelial de Aspergillus fumigatus, con el fin de utilizarlo posteriormente en pruebas diagnósticas de aspergilosis pulmonar.Para la evaluación del antígeno micelial, se emplearon técnicas de doble inmunodifusión (DD), contrainmunoelectroforesis (CIE+ ID) y enzimoinmunoensayo (ELISA), comparando sus resultados con un antígeno de referencia. La concentración de proteínas y carbohidratos del antígeno estudiado fue de 8.800pglmL y 2.452 pg/mL respectivamente, muy similares a los encontrados en el antígeno de referencia. Los antígenos analizados presentaron bandas de identidad total y parcial en la DD. No hubo bandas de precipitinas al enfrentar ambos antígenos con 20 sueros negativos, utilizando las pruebas de DD y CIE+ID. En la ClE+lD se presentaron precipitinas en los lados anódico y catódico de ambos antígenos. El punto de corte determinado en la prueba de ELlSA para el antígeno del estudio fué de 0,352, con un intervalo de confianza del 95%. Es posible concluir que el antígeno micelial estudiado puede ser empleado en pruebas serológicas diagnósticas. SummaryThis report describes the standardization and antigen characterisation of a mycelial extract of Aspergillus fumigatus, for the purpose of employing it later in pulmonary aspergillosis diagnostic tests.The mycelial antigen was evaluated using double-immunodiffusion (DD), counterimmunoelectrophoresis (CIE+ID) and enzyme immunoassay (ELISA) techniques, the results were compared with a reference antigen. The protein and carbohydrate concentrations of the mycelial antigen were 8,800 pg1mL and 2,452 pg/mL, respectively, being very similar to the reference antigen. The antigens analysed in the DD presented bands of partial and total identity. No precipitin bands were presented in DD and CIE+ID tests when both antigens with 20 negative sera were run. In the CIE+ID, precipitin bands were obsewed at the anodic and cathodic poles for both antigens. The cut-off value determined in the mycelial antigen ELISA test was 0.352 with a 95% confidence interval. It is possible to conclude that mycelial antigen can be used in diagnostic serological tests.

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Comparison of antibody measurements against Aspergillus fumigatus by means of double-diffusion and enzyme-linked immunosorbent assay (ELISA)

Cited by 69 publications

References 13 publications

Localization, Molecular Weight and Immunoglobulin Subclass Response to <i>Aspergillus fumigatus</i> Allergens in Acute Bronchopulmonary Aspergillosis

Localization, Molecular Weight and Immunoglobulin Subclass Response to <i>Aspergillus fumigatus</i> Allergens in Acute Bronchopulmonary Aspergillosis

Bronchogenic carcinoma and secondary aspergillosis—Common yet unexplored

Extracción y caracterización de antígeno micelial de Aspergillus fumigatus

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