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Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols

Legionella pneumophila are pathogenic bacteria that can be found in high concentrations in artificial water systems like evaporative cooling towers, which have been the source of frequent outbreaks in recent years. Since inhaled L. pneumophila can lead to Legionnaires’ disease, the development of su...

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Autores principales: Heining, Lena, Welp, Laura, Hugo, Achim, Elsner, Martin, Seidel, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10404198/
https://www.ncbi.nlm.nih.gov/pubmed/37204446
http://dx.doi.org/10.1007/s00216-023-04738-z
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author Heining, Lena
Welp, Laura
Hugo, Achim
Elsner, Martin
Seidel, Michael
author_facet Heining, Lena
Welp, Laura
Hugo, Achim
Elsner, Martin
Seidel, Michael
author_sort Heining, Lena
collection PubMed
description Legionella pneumophila are pathogenic bacteria that can be found in high concentrations in artificial water systems like evaporative cooling towers, which have been the source of frequent outbreaks in recent years. Since inhaled L. pneumophila can lead to Legionnaires’ disease, the development of suitable sampling and rapid analysis strategies for these bacteria in aerosols is therefore of great relevance. In this work, different concentrations of viable L. pneumophila Sg 1 were nebulized and sampled by the cyclone sampler Coriolis® µ under defined conditions in a bioaerosol chamber. To quantify intact Legionella cells, the collected bioaerosols were subsequently analyzed by immunomagnetic separation coupled with flow cytometry (IMS-FCM) on the platform rqmicro.COUNT. For analytical comparison, measurements with qPCR and cultivation were performed. Limits of detection (LOD) of 2.9 × 10(3) intact cells m(−3) for IMS-FCM and 7.8 × 10(2) intact cells m(−3) for qPCR indicating a comparable sensitivity as in culture (LOD = 1.5 × 10(3) culturable cells m(−3)). Over a working range of 10(3) − 10(6) cells mL(−1), the analysis of nebulized and collected aerosol samples with IMS-FCM and qPCR provides higher recovery rates and more consistent results than by cultivation. Overall, IMS-FCM is a suitable culture-independent method for quantification of L. pneumophila in bioaerosols and is promising for field application due to its simplicity in sample preparation. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-023-04738-z.
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spelling pubmed-104041982023-08-07 Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols Heining, Lena Welp, Laura Hugo, Achim Elsner, Martin Seidel, Michael Anal Bioanal Chem Research Paper Legionella pneumophila are pathogenic bacteria that can be found in high concentrations in artificial water systems like evaporative cooling towers, which have been the source of frequent outbreaks in recent years. Since inhaled L. pneumophila can lead to Legionnaires’ disease, the development of suitable sampling and rapid analysis strategies for these bacteria in aerosols is therefore of great relevance. In this work, different concentrations of viable L. pneumophila Sg 1 were nebulized and sampled by the cyclone sampler Coriolis® µ under defined conditions in a bioaerosol chamber. To quantify intact Legionella cells, the collected bioaerosols were subsequently analyzed by immunomagnetic separation coupled with flow cytometry (IMS-FCM) on the platform rqmicro.COUNT. For analytical comparison, measurements with qPCR and cultivation were performed. Limits of detection (LOD) of 2.9 × 10(3) intact cells m(−3) for IMS-FCM and 7.8 × 10(2) intact cells m(−3) for qPCR indicating a comparable sensitivity as in culture (LOD = 1.5 × 10(3) culturable cells m(−3)). Over a working range of 10(3) − 10(6) cells mL(−1), the analysis of nebulized and collected aerosol samples with IMS-FCM and qPCR provides higher recovery rates and more consistent results than by cultivation. Overall, IMS-FCM is a suitable culture-independent method for quantification of L. pneumophila in bioaerosols and is promising for field application due to its simplicity in sample preparation. GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00216-023-04738-z. Springer Berlin Heidelberg 2023-05-18 2023 /pmc/articles/PMC10404198/ /pubmed/37204446 http://dx.doi.org/10.1007/s00216-023-04738-z Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Paper
Heining, Lena
Welp, Laura
Hugo, Achim
Elsner, Martin
Seidel, Michael
Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title_full Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title_fullStr Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title_full_unstemmed Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title_short Immunomagnetic separation coupled with flow cytometry for the analysis of Legionella pneumophila in aerosols
title_sort immunomagnetic separation coupled with flow cytometry for the analysis of legionella pneumophila in aerosols
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10404198/
https://www.ncbi.nlm.nih.gov/pubmed/37204446
http://dx.doi.org/10.1007/s00216-023-04738-z
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