Mycopathologia

, Volume 182, Issue 3–4, pp 319–329 | Cite as

Development of a Dot-Blot Assay for the Detection of Mould-Specific IgE in the Belgian Population

  • Muriel Vincent
  • Marta Romano
  • Francis Corazza
  • Kris Huygen
  • Olivier Michel
  • Olivier Denis
Article

Abstract

Background

Data on mould sensitization in the general population are scarce and mostly on Aspergillus fumigatus, Alternaria alternata and Cladosporium herbarum.

Objectives

To validate a dot-blot assay for the detection of specific IgE and evaluate the prevalence of mould sensitization in a healthy population.

Methods

The dot-blot assay was validated against the CAP test. Sensitization rate to ten common indoor and outdoor mould species in 344 serum samples was calculated. For each serum with more than one reactivity, the “major sensitization” defined as the strongest response against a single mould species was calculated.

Results

Intra- and inter-assay variations were both below 20%, and the positivity threshold of the test was of 0.418 kU/L for A. fumigatus. Correlation with CAP results was strong. The overall prevalence of sensitization was 32.8%, and the commonest sensitizations were against A. alternaria, A. flavus and A. niger (around 15%). The most frequent “major reactivities” were against A. niger and A. alternata (20–30%). In 25.1% of the samples, “major reactivities” were directed against a group of moulds commonly found indoor (Penicillium spp., Aspergillus versicolor, Cladosporium sphaerospermum and Cladosporium cladosporioides).

Conclusions

The dot-blot assay was validated for the detection of mould-specific IgE. In the general population, sensitization to indoor species was common and accounted for 25% of overall mould sensitizations.

Keywords

IgE detection Dot-blot Mould sensitization Indoor mould species 

Abbreviation

SPT

skin prick test

Notes

Acknowledgements

We are grateful to Prof. Denis Piérard (UZ Brussel VUB) for giving us access to the serum bank from normal subjects which was used in this study and to Prof. Joël de Coninck (Laboratoire de Physique des Surfaces et des Interfaces (LPSI), Université de Mons) for his help on statistics. We thank Sofie De Prins for her excellent technical assistance. This study was supported by the Belgian Scientific Policy (BELSPO) through an Ylieff fellowship.

Authors’ Contribution

M.V, O.D and O.M designed the study protocol. M.V analysed and interpreted data and wrote the article. O.D reviewed data analysis and interpretation and revised the manuscript. O.M recruited the patients and provided critical review of the manuscript. F.C provided CAP results for allergic patients and made manuscript revision. M.R and K.H provided critical input in the manuscript framework and reviewed the paper. All authors approved the final version of the document.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no competing interests.

Supplementary material

11046_2016_91_MOESM1_ESM.docx (41 kb)
Supplementary material 1 (DOCX 41 kb)
11046_2016_91_MOESM2_ESM.docx (19 kb)
Supplementary material 2 (DOCX 19 kb)

References

  1. 1.
    Kurup VP, Shen HD, Banerjee B. Respiratory fungal allergy. Microbes Infect. 2000;2:1101–10.CrossRefPubMedGoogle Scholar
  2. 2.
    Twaroch TE, Curin M, Valenta R, Swoboda I. Mold allergens in respiratory allergy: from structure to therapy. Allergy Asthma Immunol Res. 2015;7:205–20.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Neukirch C, Henry C, Leynaert B, Liard R, Bousquet J, Neukirch F. Is sensitization to Alternaria alternata a risk factor for severe asthma? A population-based study. J Allergy Clin Immunol. 1999;103:709–11.CrossRefPubMedGoogle Scholar
  4. 4.
    Zureik M, Neukirch C, Leynaert B, Liard R, Bousquet J, Neukirch F. Sensitisation to airborne moulds and severity of asthma: cross sectional study from European community respiratory health survey. BMJ. 2002;325:411–4.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Black PN, Udy AA, Brodie SM. Sensitivity to fungal allergens is a risk factor for life-threatening asthma. Allergy. 2000;55:501–4.CrossRefPubMedGoogle Scholar
  6. 6.
    Fairs A, Agbetile J, Hargadon B, Bourne M, Monteiro WR, Brightling CE, Bradding P, Green RH, Mutalithas K, Desai D, Pavord ID, Wardlaw AJ, Pashley CH. IgE sensitization to Aspergillus fumigatus is associated with reduced lung function in asthma. Am J Respir Crit Care Med. 2010;182:1362–8.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Schwartz HJ, Citron KM, Chester EH, Kaimal J, Barlow PB, Baum GL, Schuyler MR. A comparison of the prevalence of sensitization to Aspergillus antigens among asthmatics in Cleveland and London. J Allergy Clin Immunol. 1978;62:9–14.CrossRefPubMedGoogle Scholar
  8. 8.
    Quansah R, Jaakkola MS, Hugg TT, Heikkinen SA, Jaakkola JJ. Residential dampness and molds and the risk of developing asthma: a systematic review and meta-analysis. PLoS ONE. 2012;7:e47526.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Denning DW, Pashley C, Hartl D, Wardlaw A, Godet C, Del GS, Delhaes L, Sergejeva S. Fungal allergy in asthma—state of the art and research needs. Clin Transl Allergy. 2014;4:14.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Chew GL, Rogers C, Burge HA, Muilenberg ML, Gold DR. Dustborne and airborne fungal propagules represent a different spectrum of fungi with differing relations to home characteristics. Allergy. 2003;58:13–20.CrossRefPubMedGoogle Scholar
  11. 11.
    Mahooti-Brooks N, Storey E, Yang C, Simcox NJ, Turner W, Hodgson M. Characterization of mold and moisture indicators in the home. J Occup Environ Hyg. 2004;1:826–39.CrossRefPubMedGoogle Scholar
  12. 12.
    Bellanger AP, Reboux G, Roussel S, Grenouillet F, Didier-Scherer E, Dalphin JC, Millon L. Indoor fungal contamination of moisture-damaged and allergic patient housing analysed using real-time PCR. Lett Appl Microbiol. 2009;49:260–6.CrossRefPubMedGoogle Scholar
  13. 13.
    Alshareef F, Robson GD. Prevalence, persistence, and phenotypic variation of Aspergillus fumigatus in the outdoor environment in Manchester, UK, over a 2-year period. Med Mycol. 2014;52:367–75.CrossRefPubMedGoogle Scholar
  14. 14.
    Beguin H. Mould biodiversity in homes I. Air and surfaces analysis of 130 dwellings. Aerobiologia. 1994;10:157–66.CrossRefGoogle Scholar
  15. 15.
    Reboux G, Bellanger AP, Roussel S, Grenouillet F, Sornin S, Piarroux R, Dalphin JC, Millon L. Indoor mold concentration in Eastern France. Indoor Air. 2009;19:446–53.CrossRefPubMedGoogle Scholar
  16. 16.
    Achatz G, Oberkofler H, Lechenauer E, Simon B, Unger A, Kandler D, Ebner C, Prillinger H, Kraft D, Breitenbach M. Molecular cloning of major and minor allergens of Alternaria alternata and Cladosporium herbarum. Mol Immunol. 1995;32:213–27.CrossRefPubMedGoogle Scholar
  17. 17.
    Huygen K, Rodeghiero C, Govaerts D, Leroux-Roels I, Melin P, Reynders M, Van Der Meeren S, Van Den Wijngaert S, Pierard D. Bordetella pertussis seroprevalence in Belgian adults aged 20–39 years, 2012. Epidemiol Infect. 2014;142:724–8.CrossRefPubMedGoogle Scholar
  18. 18.
    Soeria-Atmadja D, Onell A, Borga A. IgE sensitization to fungi mirrors fungal phylogenetic systematics. J Allergy Clin Immunol. 2010;125:1379–86.CrossRefPubMedGoogle Scholar
  19. 19.
    Bousquet PJ, Chinn S, Janson C, Kogevinas M, Burney P, Jarvis D. Geographical variation in the prevalence of positive skin tests to environmental aeroallergens in the European community respiratory health survey I. Allergy. 2007;62:301–9.CrossRefPubMedGoogle Scholar
  20. 20.
    Zock JP, Jarvis D, Luczynska C, Sunyer J, Burney P. Housing characteristics, reported mold exposure, and asthma in the European community respiratory health survey. J Allergy Clin Immunol. 2002;110:285–92.CrossRefPubMedGoogle Scholar
  21. 21.
    Arshad SH, Tariq SM, Matthews S, Hakim E. Sensitization to common allergens and its association with allergic disorders at age 4 years: a whole population birth cohort study. Pediatrics. 2001;108:E33.CrossRefPubMedGoogle Scholar
  22. 22.
    Arbes SJ Jr, Gergen PJ, Elliott L, Zeldin DC. Prevalences of positive skin test responses to 10 common allergens in the US population: results from the third National Health and Nutrition Examination Survey. J Allergy Clin Immunol. 2005;116:377–83.CrossRefPubMedGoogle Scholar
  23. 23.
    Sears MR, Herbison GP, Holdaway MD, Hewitt CJ, Flannery EM, Silva PA. The relative risks of sensitivity to grass pollen, house dust mite and cat dander in the development of childhood asthma. Clin Exp Allergy. 1989;19:419–24.CrossRefPubMedGoogle Scholar
  24. 24.
    Mari A, Schneider P, Wally V, Breitenbach M, Simon-Nobbe B. Sensitization to fungi: epidemiology, comparative skin tests, and IgE reactivity of fungal extracts. Clin Exp Allergy. 2003;33:1429–38.CrossRefPubMedGoogle Scholar
  25. 25.
    Hemmann S, Blaser K, Crameri R. Allergens of Aspergillus fumigatus and Candida boidinii share IgE-binding epitopes. Am J Respir Crit Care Med. 1997;156:1956–62.CrossRefPubMedGoogle Scholar
  26. 26.
    Crameri R, Zeller S, Glaser AG, Vilhelmsson M, Rhyner C. Cross-reactivity among fungal allergens: a clinically relevant phenomenon? Mycoses. 2009;52:99–106.CrossRefPubMedGoogle Scholar
  27. 27.
    O’Hollaren MT, Yunginger JW, Offord KP, Somers MJ, O’Connell EJ, Ballard DJ, Sachs MI. Exposure to an aeroallergen as a possible precipitating factor in respiratory arrest in young patients with asthma. N Engl J Med. 1991;324:359–63.CrossRefPubMedGoogle Scholar
  28. 28.
    Baur X, Sander I, Posch A, Raulf-Heimsoth M. Baker’s asthma due to the enzyme xylanase–a new occupational allergen. Clin Exp Allergy. 1998;28:1591–3.CrossRefPubMedGoogle Scholar
  29. 29.
    Wang DY, Hadj-Henni L, Thierry S, Arne P, Chermette R, Botterel F, Hadrich I, Makni F, Ayadi A, Ranque S, Huang WY, Guillot J. Simple and highly discriminatory VNTR-based multiplex PCR for tracing sources of Aspergillus flavus isolates. PLoS ONE. 2012;7:e44204.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Krishnan S, Manavathu EK, Chandrasekar PH. Aspergillus flavus: an emerging non-fumigatus Aspergillus species of significance. Mycoses. 2009;52:206–22.CrossRefPubMedGoogle Scholar
  31. 31.
    Hedayati MT, Pasqualotto AC, Warn PA, Bowyer P, Denning DW. Aspergillus flavus: human pathogen, allergen and mycotoxin producer. Microbiology. 2007;153:1677–92.CrossRefPubMedGoogle Scholar
  32. 32.
    Vermani M, Vijayan VK, Agarwal MK. Identification of Aspergillus (A. flavus and A. niger) Allergens and Heterogeneity of Allergic Patients’ IgE Response. Iran J Allergy Asthma Immunol. 2015;14:361–9.PubMedGoogle Scholar
  33. 33.
    Karvonen AM, Hyvarinen A, Korppi M, Haverinen-Shaughnessy U, Renz H, Pfefferle PI, Remes S, Genuneit J, Pekkanen J. Moisture damage and asthma: a birth cohort study. Pediatrics. 2015;135:e598–606.CrossRefPubMedGoogle Scholar
  34. 34.
    Fisk WJ, Lei-Gomez Q, Mendell MJ. Meta-analyses of the associations of respiratory health effects with dampness and mold in homes. Indoor Air. 2007;17:284–96.CrossRefPubMedGoogle Scholar
  35. 35.
    Iossifova YY, Reponen T, Ryan PH, Levin L, Bernstein DI, Lockey JE, Hershey GK, Villareal M, LeMasters G. Mold exposure during infancy as a predictor of potential asthma development. Ann Allergy Asthma Immunol. 2009;102:131–7.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Muriel Vincent
    • 1
  • Marta Romano
    • 2
  • Francis Corazza
    • 3
  • Kris Huygen
    • 1
    • 2
  • Olivier Michel
    • 4
  • Olivier Denis
    • 1
  1. 1.Program Allergology - Scientific Service ImmunologyWIV-ISP (site Uccle)BrusselsBelgium
  2. 2.Program Host-Pathogen Interaction - Scientific Service ImmunologyWIV-ISP (site Uccle)BrusselsBelgium
  3. 3.Laboratory of Immunology and of Translational ResearchCHU Brugmann (Université Libre de Bruxelles - ULB)BrusselsBelgium
  4. 4.Clinic of Immunology and AllergologyCHU Brugmann (Université Libre de Bruxelles - ULB)BrusselsBelgium

Personalised recommendations