caa a22 4500 60550279X CHVBK 20210128100607.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6785-9 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6785-9 Development and performance assessment of a qualitative SYBR® green real-time PCR assay for the detection of Aspergillus versicolor in indoor air [Elektronische Daten] [X. Libert, C. Chasseur, S. Bladt, A. Packeu, F. Bureau, N. Roosens, S. De Keersmaecker] Currently, contamination of indoor environment by fungi and molds is considered as a public health problem. The monitoring of indoor airborne fungal contamination is a common tool to help understanding the link between fungi in houses and respiratory problems. Classical analytical monitoring methods, based on cultivation and microscopic identification, depend on the growth of the fungi. Consequently, they are biased by difficulties to grow some species on certain culture media and under certain conditions or by noncultivable or dead fungi that can consequently not be identified. However, they could have an impact on human health as they might be allergenic. Since molecular methods do not require a culture step, they seem an excellent alternative for the monitoring of indoor fungal contaminations. As a case study, we developed a SYBR® green real-time PCR-based assay for the specific detection and identification of Aspergillus versicolor, which is frequently observed in indoor environment and known to be allergenic. The developed primers amplify a short region of the internal transcribed spacer 1 from the 18S ribosomal DNA complex. Subsequently, the performance of this quantitative polymerase chain reaction (qPCR) method was assessed using specific criteria, including an evaluation of the selectivity, PCR efficiency, dynamic range, and repeatability. The limit of detection was determined to be 1 or 2 copies of genomic DNA of A. versicolor. In order to demonstrate that this SYBR® green qPCR assay is a valuable alternative for monitoring indoor fungal contamination with A. versicolor, environmental samples collected in contaminated houses were analyzed and the results were compared to the ones obtained with the traditional methods. The Author(s), 2015 Aspergillus versicolor nationallicence Indoor air nationallicence Public health nationallicence Real-time PCR nationallicence SYBR® green nationallicence Performance assessment nationallicence Libert X. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut Chasseur C. Health and Environment, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut Bladt S. Cellule Régionale d'Intervention en Pollution Intérieure (CRIPI), Brussels Environment (IBGE), Avenue du Port 86C/3000, 1000, Bruxelles, Belgium aut Packeu A. Mycology and Aerobiology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut Bureau F. Cellular and Molecular Immunology, Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA), Université de Liège (ULg), Avenue de l'Hôpital, 1 (B34), 4000, Sart-Tilman, Belgium aut Roosens N. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut De Keersmaecker S. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7267-7282 0175-7598 99:17<7267 2015 99 253 https://doi.org/10.1007/s00253-015-6785-9 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6785-9 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Libert X. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Chasseur C. Health and Environment, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Bladt S. Cellule Régionale d'Intervention en Pollution Intérieure (CRIPI), Brussels Environment (IBGE), Avenue du Port 86C/3000, 1000, Bruxelles, Belgium aut NATIONALLICENCE 700 1- Packeu A. Mycology and Aerobiology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Bureau F. Cellular and Molecular Immunology, Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA), Université de Liège (ULg), Avenue de l'Hôpital, 1 (B34), 4000, Sart-Tilman, Belgium aut NATIONALLICENCE 700 1- Roosens N. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- De Keersmaecker S. Platform Biotechnology and Molecular Biology, Scientific Institute of Public Health (WIV-ISP), J. Wytsmanstraat 14, 1050, Brussels, Belgium aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7267-7282 0175-7598 99:17<7267 2015 99 253