Aspergillus Fumigatus ELISA Kit

Intended Use:
The Bordier Aspergillus fumigatus ELISA kit is intended for the quantitative detection of IgG antibodies against Aspergillus fumigatus in human serum. Serology is an aid for diagnosis and cannot be used as the sole method of diagnosis. This test is also intended for follow-up of patients at risk for aspergillosis infections.

Background:
Pulmonary aspergillosis is caused by different pathogenic species of the fungal genus Aspergillus, the most frequently implicated being Aspergillus fumigatus. This pathogen is found in soil and decaying organic matter. Humans inhale hundreds of spores per day, but only people with risk factors will develop different types of aspergillosis: allergic bronchopulmonary aspergillosis, allergic sinusitis, aspergilloma and chronic pulmonary aspergillosis. The main symptoms are couch and shortness of breath. Since theses symptoms are unspecific, diagnosis is based on a combination of clinical, radiological, biological, and mycological criteria. Serology is an important criterion and several methods are available for screening, follow-up and confirmation.

Principle and Presentation:
The kit provides all the material needed to perform 96 enzyme-linked immunosorbent assays (ELISA) on breakable microtitration wells sensitized with the following mix:

– Solubles somatic and metabolic Aspergillus fumigatus antigens
– Recombinants antigens: dipeptidylpeptidase type V (chymotrypsin) and ribonuclease (mitogillin) from Aspergillus fumigatus

Specific antibodies in the sample will bind to these antigens and washing will remove unspecific antibodies. The presence of fungal specific antibodies is detected with a Protein A – alkaline phosphatase conjugate. A second washing step will remove unbound conjugate. Revealing bound antibodies is made by the addition of pNPP substrate which turns yellow in the presence of alkaline phosphatase. Color intensity is proportional to the amount of Aspergillus fumigatus specific antibodies in the sample. Potassium phosphate is added to stop the reaction. Absorbance at 405 nm is read using an ELISA microplate reader.

The test can be performed with automatic systems, but this must be validated by the user.