Description:

1. Intended UseThe GenWay Aspergillus fumigatus IgA Antibody ELISA Test Kit has been designed for the thedetection and the quantitative determination of specific IgA antibodies against Aspergillusfumigatus in serum and plasma. Further applications in other body fluids are possible and can berequested from the Technical Service of GenWay .This assay is intended for research use only.

2. General InformationAspergillus species of known pathogenicity to man are Aspergillus fumigatus, A. flavus, A. niger, A.terreus and A. nidulans. The most common pathogen of this genus is A. fumingatus which occurs inhay, grain, rotten plants and birds faeces. The main opportunistic invasive fungial infections are thecandidal mycosis followed by aspergillosis. Generally infections with Aspergillus spp. are airborne.Because of the ubiquity of Aspergillus species it renders more difficult to decide betweencontamination by commensals or a serious infection. Usually infection in man occurs in alreadydamaged tissues only. Aspergillus spp. can cause a chronical infection of paranasal sinus, eyes orlungs.Three types of lung-aspergillosis can be distinguished:a: Acute infection (bronchial pneumonia; pneumonia)Aspergillus pneumonia is mostly found in individuals with neutropenia (decrease of neutrophilgranulocytes), after a long-time therapy with glucocorticoids, in immunosuppressed individuals(after organ transplantation) and in alcoholics.b: Saprophytic aspergillom (compact reticulum of hyphae in the lungs)Preformed caves in the lung due to a previous tuberculosis give place to a colonisation ofAspergillus species.c: Allergic bronchopulmonal aspergillosisThis clinical picture is not due to an infectious disease but a hypersensitive reaction of the bronchialsystem (mediated by IgE) after inhalation of aspergillus spores. Subsequently the bronchial systemproduces highly viscous secretions, that may block the bronchial lumen. The individual developsdifficulties of breathing and a fibrosis.Next to ELISA the indirect Aspergillus hemagglutination test (Aspergillus HAT) can be performedto detect specific IgG and IgM antibodies. The HAT is not suitable as a screening test, however,because of its low sensitivity. In some high-risk individuals it shows only low antibody titers. For abetter diagnosis of invasive aspergillosis the brain or lung of these individuals should be examinedby a biopsy.

3. Principle of the TestThe GenWay Aspergillus fumigatus IgA antibody test kit is based on the principle of the enzymeimmunoassay (EIA). Aspergillus antigen is bound on the surface of the microtiter strips. Dilutedsample serum or ready-to-use standards are pipetted into the wells of the microtiter plate. A bindingbetween the IgA antibodies of the serum and the immobilized Aspergillus antigen takes place. Aftera one hour incubation at room temperature, the plate is rinsed with diluted wash solution, in order toremove unbound material. Then ready-to-use anti-human-IgA peroxidase conjugate is added andincubated for 30 minutes. After a further washing step, the substrate (TMB) solution is pipetted andincubated for 20 minutes, inducing the development of a blue dye in the wells. The colordevelopment is terminated by the addition of a stop solution, which changes the color from blue toyellow. The resulting dye is measured spectrophotometrically at the wavelength of 450 nm. Theconcentration of the IgA antibodies is directly proportional to the intensity of the color.

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