Analytical bias of cross-reactive polyclonal antibodies for environmental immunoassays of Alternaria alternata

J Allergy Clin Immunol. 2008 Mar;121(3):763-8. doi: 10.1016/j.jaci.2007.09.046. Epub 2007 Nov 26.

Abstract

Background: Alternaria alternata is recognized as an important aeroallergen indoors and outdoors, and exposure to the fungus has been identified as a risk factor for asthma. Two recent publications concluded that 95% to 99% of American homes contained detectable amounts of Alternaria antigens when analyzed with a polyclonal antibody (pAb)-based ELISA.

Objectives: We investigated the cross-reactivity of the commercially available pAbs that were used in those studies.

Methods: Reactivity to 24 fungal species commonly found in indoor environments was analyzed by inhibition ELISA by using solid-phase A alternata antigen. The pAbs were also tested by immunoblotting and halogen immunoassay for a subgroup of fungi.

Results: Spores of 7 fungi including species of Alternaria, Ulocladium, Stemphylium, Epicoccum, Drechslera, and Exserohilum strongly inhibited the binding of the pAbs when tested by ELISA. Six other fungi reacted in the ELISA at a lower level, and 11 fungal species including several Penicillium, Aspergillus, Fusarium, and Cladosporium species failed to show inhibition. The immunoblots and the halogen immunoassay staining confirmed the cross-reactivity patterns of the ELISA.

Conclusion: The pAbs against A alternata were found to cross-react broadly with related and nonrelated fungi. The prevalence data previously reported for A alternata should be considered to be fungal-reactive rather than A alternata-specific.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Air Microbiology*
  • Air Pollution, Indoor
  • Alternaria / isolation & purification*
  • Antibodies, Fungal / immunology*
  • Antibody Specificity
  • Antigens, Fungal / analysis*
  • Cross Reactions
  • Environmental Monitoring / methods*
  • Immunoassay* / methods
  • Immunoblotting

Substances

  • Antibodies, Fungal
  • Antigens, Fungal