Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy

• DOI: 10.1128/aem.00929-15
• Publication Date: 2015-06-20T02:57:08Z
• Authors: Komlavi Anani Afanou; Anne Straumfors; Asbjørn Skogstad; Ajay P. Nayak; Ida Skaar; Linda Hjeljord; Arne Tronsmo; Wijnand Eduard; Brett James Green
• Keywords: Immunolabeling; Polyclonal antibodies

Submicronic fungal fragments have been observed in vitro aerosolization experiments. The occurrence of these particles has therefore suggested to contribute respiratory health problems mold-contaminated indoor environments. However, the role submicronic exacerbating adverse effects remained unclear due limitations associated with detection methods. In present study, we report development an indirect immunodetection assay that utilizes chicken polyclonal antibodies developed against spores from Aspergillus versicolor and high-resolution field emission scanning electron microscopy (FESEM). Immunolabeling was performed A. immobilized fixed onto poly-l-lysine-coated polycarbonate filters. Ninety percent 1- 2-μm fragments, compared 100% >2-μm generated pure freeze-dried mycelial versicolor, were positively labeled. proof-of-concept experiments, air samples collected moldy environments evaluated using immunolabeling technique. Our results indicated 13% total derived fungi. This fraction comprises 79% detected by 21% spore morphologically identified. methods reported this study enable enumeration particles, including a complex heterogeneous environmental sample.