Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy.

Abstract:

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

journal_name

Appl Environ Microbiol

authors

Afanou KA,Straumfors A,Skogstad A,Nayak AP,Skaar I,Hjeljord L,Tronsmo A,Eduard W,Green BJ

doi

10.1128/AEM.00929-15

subject

Has Abstract

pub_date

2015-09-01 00:00:00

pages

5794-803

issue

17

eissn

0099-2240

issn

1098-5336

pii

AEM.00929-15

journal_volume

81

pub_type

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