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Publikationstyp
Conference proceedings
Conference proceedings
Konferenzposter
Forschungsbericht
Monographie
Conference proceedings
Konferenzposter
Forschungsbericht
Monographie
Erscheinungsjahr
2015
DOI
Mould Specific Quantitative PCR
Mould Specific Quantitative PCR
Evaluation of the applicability of a Mould Specific Quantitative PCR (MSQPCR) assay for mould detection in Central European indoor environments
Autor:innen
Herausgeber
Quelle
49. Wissenschaftliche Tagung der Deutschsprachigen Mykologischen Gesellschaft e.V. (49, 2015, Jena)
1st International Symposium of the CRC/Transregio FungiNet/ DMykG (1, 2015 Jena)
1st International Symposium of the CRC/Transregio FungiNet/ DMykG (1, 2015 Jena)
Schlagwörter
Finanzierungskennzeichen
371161236
standardisiertes Finanzierungskennzeichen
37116123
Verbundene Publikation
Zitation
Mould Specific Quantitative PCR, 2015. [online]. Verfügbar unter: https://openumwelt.de/handle/123456789/7665
Zusammenfassung englisch
Spores and conidia of various filamentous fungi are a potential health risk in indoor environments. Currently, convenient, rapid and specific PCR methods for detection of spores and conidia are missing. The MSQPCR assay offers a putative solution allowing the detection of 36 different mould fungi. However, it is not known whether the species selectivity of MSQPCR is sufficient for German indoor environments and how high the ability of the assay is to discriminate between the target species.
We evaluated 108 MSQPCR primers for their specificity, binding affinity, genome locality, oligo quality and PCR product size with NCBI-Genbank Primer BLAST (National Centre of Biotechnology Information, Bethesda), Primer 3 (Untergrasser et al. 2007) and Oligocalc (Kibbe, 2007).. Fungal species frequently detected in water and mould damaged buildings in Germany were compared with the species selection of MSQPCR system to evaluate its applicability for German indoor environments.
Stachybotrys chartarum, Aspergillus versicolor and Penicillum chrysogenum were used for practical testing of MSQPCR assay. For this purpose, the three reference strains, 5 to 10 additional isolates of the target species and 20 non target species were tested. DNA was extracted from a filtered spore dilution and subjected to the MSQPCR assay. From 108 primers tested, 13% and 24% had no homologies to other than target species on genus level and beyond genus level, respectively. This result was confirmed by the conducted spore experiment with spores of multiple strains.
This work demonstrates that most of the fungal species detected with MSQPCR assay are relevant for European indoor water and mould damages. However, the species spectra should be enlarged and adapted to European conditions. The MSQPCR qPCR primers were not considered specific enough for the detection of target species. We suggest to modify the test system by improved specific primers.
© Valtanen K, Baschien C., Jacobs K., Szwezyk, R.
We evaluated 108 MSQPCR primers for their specificity, binding affinity, genome locality, oligo quality and PCR product size with NCBI-Genbank Primer BLAST (National Centre of Biotechnology Information, Bethesda), Primer 3 (Untergrasser et al. 2007) and Oligocalc (Kibbe, 2007).. Fungal species frequently detected in water and mould damaged buildings in Germany were compared with the species selection of MSQPCR system to evaluate its applicability for German indoor environments.
Stachybotrys chartarum, Aspergillus versicolor and Penicillum chrysogenum were used for practical testing of MSQPCR assay. For this purpose, the three reference strains, 5 to 10 additional isolates of the target species and 20 non target species were tested. DNA was extracted from a filtered spore dilution and subjected to the MSQPCR assay. From 108 primers tested, 13% and 24% had no homologies to other than target species on genus level and beyond genus level, respectively. This result was confirmed by the conducted spore experiment with spores of multiple strains.
This work demonstrates that most of the fungal species detected with MSQPCR assay are relevant for European indoor water and mould damages. However, the species spectra should be enlarged and adapted to European conditions. The MSQPCR qPCR primers were not considered specific enough for the detection of target species. We suggest to modify the test system by improved specific primers.
© Valtanen K, Baschien C., Jacobs K., Szwezyk, R.