Quantitation of Aspergillus versicolor Antigen Using ELISA and Fluorescent Multiplex Array Technology

B.R. Smith, S. Kespohl, I. Sander, M. Raulf-Heimsoth, E.M. King, M.D. Chapman

Author address: 

INDOOR Biotechnologies, Charlottesville, VA, Institue for Prevention and Occupational Medicine of the German Social Accident Insurance, Ruhr University, Bochum, GERMANY.

Abstract: 

RATIONALE: Aspergillus versicolor is an allergenic and potentially toxigenic mold found in damp indoor environments and water-damaged buildings. The health implications associated with mold exposure are not well understood. Reliable methods for exposure assessment are required to determine associated risks. Our aim was to develop sensitive immunoassays for the detection of A. versicolor antigen. METHODS: Five monoclonal anti-A. versicolor antibodies were used in ELISA to screen extracts of 10 different mold species commonly found in water-damaged buildings. The extracts were also tested by fluorescent multiplex array using monoclonal antibodies covalently coupled to the surface of internally labeled microspheres. Extracts prepared from mold-contaminated wallboard material were tested by mAb ELISA as well as by polyclonal ELISA for A. versicolor antigen. RESULTS: The ELISA and multiplex array were shown to be specific for A. versicolor and recognized an antigen designated AveX. Both assays exhibited high levels of sensitivity: dynamic ranges were 1000-1.9ng/ml for ELISA and 500-0.24ng/ml for the fluorescent multiplex array. The polyclonal ELISA had a dynamic range of 10-0.04ng/mL. Measurements of AveX in wallboard samples (n=11) correlated with the polyclonal ELISA, particularly for samples with relatively high antigen content. CONCLUSIONS: We have developed species-specific and highly sensitive assays for Aspergillus versicolor antigen using monoclonal antibody-based ELISA and fluorescent multiplex array. The data suggest that murine antibody responses to A. versicolor tend to be species specific and/or that A.versicolor produces unique antigens. These assays may be useful for A.versicolor exposure assessment and to determine associated health risks.
2011

abstract No: 

AB168

Full conference title: 

American Academy of Allergy Asthma & Immunology
    • AAAAI 2011 (67th)