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Validation of a quantitative PCR based detection system for indoor mold exposure assessment in bioaerosols

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Abstract

Determination and assessment of airborne fungal particles is complex and results of different sampling and analytical strategies are hard to compare due to limitations of each of the techniques. Here, an indoor mold detection system based on quantitative polymerase chain reaction (qPCR) is described and validated for its reliability and stability to identify airborne fungal particles collected. Data obtained from testing the system with fungal DNA, spore suspensions and bioaerosols indicated a need for spiking and normalization of measurements due to material loss and assay specific bias. Considering the loss of material during sample processing, detection limits defined for suspensions of Tritirachium oryzae spores were roughly 18 spores per sample. Detection of fungal spore mixtures nebulized under controlled conditions in a bioaerosol chamber showed generally 2–3 times higher normalized values measured with the molecular system compared to cultivation. Data obtained from a mold infested indoor sampling site and its corresponding outdoor reference measurement showed good correlations between qPCR and high-throughput sequencing (rho = 0.83, p < 0.01), if Cladosporium species were excluded. Taking necessary data normalization into account, the described qPCR detection system shows great potential to complement commonly used culture based approaches with the aim to improve the precision of indoor mold assessments. In contrast to already available qPCR assays that detect certain molds on a species level, this system covers a broad range of relevant fungal communities, serving as a promising alternative to high-throughput sequencing to identify indoor molds.

Graphical abstract: Validation of a quantitative PCR based detection system for indoor mold exposure assessment in bioaerosols

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Publication details

The article was received on 05 Jun 2018, accepted on 05 Sep 2018 and first published on 06 Sep 2018


Article type: Paper
DOI: 10.1039/C8EM00253C
Citation: Environ. Sci.: Processes Impacts, 2018, Advance Article
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    Validation of a quantitative PCR based detection system for indoor mold exposure assessment in bioaerosols

    V. Unterwurzacher, C. Pogner, H. Berger, J. Strauss, S. Strauss-Goller and M. Gorfer, Environ. Sci.: Processes Impacts, 2018, Advance Article , DOI: 10.1039/C8EM00253C

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