Analytical bias of cross-reactive polyclonal antibodies for environmental immunoassays of Alternaria alternata - 15/08/11
, Brett J. Green, PhD, Francoise M. Blachere, MSc, Erika Janotka, MUDr, Donald H. Beezhold, PhD
Reprint requests: Detlef Schmechel, PhD, Allergy and Clinical Immunology Branch, National Institute for Occupational Safety and Health, 1095 Willowdale Road, Morgantown, WV 26505.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.
Le texte complet de cet article est disponible en PDF.Key words : Fungi, polyclonal antibodies, Alternaria, ELISA
Abbreviations used : HIA, pAb, PBST, PBSTM, RT
Plan
| Supported in part by an Interagency Agreement from the National Institute of Environmental Health Sciences (Y1-ES-0001)-Immunotoxicity of Workplace Xenobiotics and with internal National Institute for Occupational Safety and Health funding. |
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| The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the National Institute for Occupational Safety and Health. |
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| Disclosure of potential conflict of interest: The authors have declared that they have no conflict of interest. |
Vol 121 - N° 3
P. 763-768 - mars 2008 Retour au numéro
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