Quantification of the Aspergillus versicolor allergen in house dust

Shi, Chunhua; Belisle, Donald; Miller, J. David

doi:10.1016/j.jim.2011.06.034

Cited by 14 publications

(4 citation statements)

References 23 publications

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“…Similar results have been reported with polyclonal antibodies (pAbs) raised against spores from A. versicolor (49)(50)(51) or other fungal species (52) where high levels of cross-reactivity were observed. Even monoclonal antibodies developed after immunization of animals with spores/conidia were found not to be species specific (49,53).…”

Section: Discussionsupporting

confidence: 86%

Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy

Afanou

Straumfors

Skogstad

et al. 2015

Appl Environ Microbiol

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dSubmicronic 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-m fragments, compared to 100% of >2-m 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. P ersonal exposure to fungal aerosols in damp buildings has been associated with respiratory morbidity (1, 2). Fungal spore exposure levels in residential indoor environments seem too low to explain such an association (3). Submicronic fungal fragments observed in vitro during aerosolization experiments with fungal cultures are believed to contribute to the respiratory health problems observed in moldy indoor environments (4, 5). However, this role of submicronic fragments has remained unclear due to limitations associated with their quantification.Airborne fungal particles have been shown to include spores in addition to larger and smaller (submicronic) fragments of spores and hyphae. These fragments may constitute a significant reservoir for antigens, allergens, and toxins in addition to spores. To date, the quantification of submicronic fungal fragments has remained technically challenging in environmental samples due to the lack of adequate detection and enumeration methods (6, 7). In this regard, the evaluation of the exposure burden of fungal submicronic fragments in fungally contaminated environments has been underestimated. In vitro studies that have evaluated the release of submicronic fragments have provided insight into the aerodynamic characteristics as well as the abiotic factors that influence the release of these particles. These laboratory studies of common indoor fungal isolates have shown the need to include the enumeration of submicronic fragments in addition to spores and larger fragments during expos...

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Section: Discussionsupporting

confidence: 86%

Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy

Afanou

Straumfors

Skogstad

et al. 2015

Appl Environ Microbiol

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“…versicolor has been reported. 38 This sandwich ELISA is based on polyclonal antibodies against the single allergen Asp v 13, which is characteristic of A. versicolor spores. The assay has a detection limit of 24 pg ml À1 and is ve-fold more sensitive than our assay.…”

Section: Discussionmentioning

confidence: 99%

A new immunoassay to quantify fungal antigens from the indoor mould Aspergillus versicolor

Zahradnik

Kespohl

Sander

et al. 2013

Environ. Sci.: Processes Impacts

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Aspergillus versicolor is among the most commonly found moulds in moisture-damaged buildings and can be associated with adverse health effects in humans. This paper reports the development, validation and application of an enzyme immunoassay to quantify A. versicolor antigens. A sandwich ELISA was developed using polyclonal antibodies that recognize a broad range of A. versicolor proteins present in fungal spores and in mycelia fragments. To validate the new method, A. versicolor antigens were quantified in samples collected from homes with visible mould growth, including dust from vacuumed walls and bulk samples of building materials. Antigen concentrations were compared to the results of a commercial ELISA based on monoclonal antibodies (AveX ELISA, Indoor Biotechnologies, Charlottesville, USA) and correlated with colony forming units (CFU) of A. versicolor. The A. versicolor ELISA was very sensitive with a lower detection limit of 120 pg ml(-1). The assay also showed some reactivity to other moulds with strongest reactions with other Aspergillus species (1-3% reactivity). The new assay detected A. versicolor antigens in a much higher percentage of dust samples (88% vs. 27%) and bulk samples (89% vs. 24%) than the AveX assay. A significant correlation (r = 0.67, and p < 0.0001) was found between antigen concentrations and CFU of A. versicolor. Based on its low detection limit and good correlation with the culture-based method, this new immunoassay seems to be a useful tool for the measurement of A. versicolor exposure levels and a reliable complement to the traditional monitoring techniques, such as mould cultivation or microscopy.

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“…68 Porous materials that allow absorption and accumulation of dust are generally excellent substrates for the growth of Aspergillus versicolor . 77 The species Aspergillus versicolor is one of the most aggressive species to health, with known allergenic and pathogenic characteristics. 78 Furthermore, both Penicillium and Aspergillus possess species that produce mycotoxins and may be responsible for some of the symptoms associated to sick building syndrome.…”

Section: Fungimentioning

confidence: 99%

Indoor air quality in an Antarctic Research Station: Fungi, particles and aldehyde concentrations associated with building materials and architectural design

Pagel

Reis

Alvarez

et al. 2017

Indoor and Built Environment

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Antarctic buildings are enclosed structures, which provide shelter and logistic support to researchers and personnel who remain indoors for long periods and can be affected by air pollution caused by construction materials and activities inside buildings. This study aims to investigate the indoor air quality at the Brazilian Comandante Ferraz Antarctic Station based on measurements of aldehydes, particulate matter and fungi conducted during the Antarctic summer in 2012. The sampling site was divided in conditioned (personnel living quarters) and unconditioned (service and utilities areas) compartments and outdoor sites. A field log book was used to record the activities in the station. Furniture and plywood coverings may have contributed to high average concentrations of formaldehyde. Cooking resulted in high average levels of acrolein and fine particles in most of the monitored environments. Other activities such as cleaning, use of personal and cosmetic products, waste incineration, building maintenance and movement of people and vehicles have also contributed to particles concentration increase. Dominance of the species Aspergillus versicolor and Penicillium sp. showed potential means of fungal proliferation. Considering that the functionality and operation are similar in many Antarctic buildings, some general recommendations were outlined.

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Quantification of the Aspergillus versicolor allergen in house dust

Cited by 14 publications

References 23 publications

Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy

Indirect Immunodetection of Fungal Fragments by Field Emission Scanning Electron Microscopy

A new immunoassay to quantify fungal antigens from the indoor mould Aspergillus versicolor

Indoor air quality in an Antarctic Research Station: Fungi, particles and aldehyde concentrations associated with building materials and architectural design

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