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Applied Microbiology
Volume 4
Issue 1
10.3390/applmicrobiol4010019
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Article

Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry

by
Philipp Streich
1,
Johannes Redwitz
2,
Sandra Walser-Reichenbach
2,
Caroline E. W. Herr
2,3,
Martin Elsner
1 and
Michael Seidel
1,*
1
Chair of Analytical Chemistry and Water Chemistry, TUM School of Natural Sciences, Technical Universtity of Munich, Lichtenbergstraße 4, 85748 Garching, Germany
2
Department of Occupational and Environmental Health, Epidemiology, Bavarian Health and Food Safety Authority, Pfarrstraße 3, 80538 Munich, Germany
3
Institute and Clinic for Occupational, Social and Environmental Medicine, University Hospital, LMU Munich, Pettenkoferstraße 8a, 80336 Munich, Germany
*
Author to whom correspondence should be addressed.
Appl. Microbiol. 2024, 4(1), 284-296; https://doi.org/10.3390/applmicrobiol4010019
Submission received: 18 December 2023 / Revised: 22 January 2024 / Accepted: 24 January 2024 / Published: 27 January 2024
(This article belongs to the Special Issue Exclusive Papers Collection of Editorial Board Members and Invited Scholars in Applied Microbiology (2023, 2024))
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Abstract

Legionella pneumophila are pathogenic bacteria that repeatedly occur in high concentrations in the process water of evaporative cooling systems (ECS). When released into the environment, the resulting bioaerosols can cause outbreaks with fatal consequences. The official, internationally accepted detection method for Legionella spp. in water samples is based on cultivation. However, cultivation is time-consuming and may underestimate the total count of viable L. pneumophila in ECS. Therefore, culture-independent methods are receiving attention for rapid monitoring. Cartridge-based immunomagnetic separation (IMS) coupled with flow cytometry (FCM) is an innovative, antibody-based method for the culture-independent quantification of L. pneumophila, using a panel of antibodies against serogroup (Sg) 1–15. We characterized the IMS-FCM method as a quantitative rapid test by general analytical procedures. Viable cryopreserved L. pneumophila standards were used in calibration experiments for the method. We achieved detection limits for Sg 1, Sg 4, and Sg 6 of 100, 105 and 88 viable cells per 100 mL, respectively. Furthermore, we demonstrated the practical applicability of IMS-FCM with real ECS samples and compared the performance against cultivation. Cultivation showed here no positive results, but IMS-FCM evidenced L. pneumophila in a range of 0–80,000 viable cells per 100 mL. This work demonstrates that IMS-FCM is a suitable, culture-independent, quantitative method for rapidly monitoring L. pneumophila.
Keywords: Legionella pneumophila; process water; culture-independent; quantitative rapid test; viable cryopreserved bacteria standards; immunomagnetic separation; flow cytometry Legionella pneumophila; process water; culture-independent; quantitative rapid test; viable cryopreserved bacteria standards; immunomagnetic separation; flow cytometry

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MDPI and ACS Style

Streich, P.; Redwitz, J.; Walser-Reichenbach, S.; Herr, C.E.W.; Elsner, M.; Seidel, M. Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry. Appl. Microbiol. 2024, 4, 284-296. https://doi.org/10.3390/applmicrobiol4010019

AMA Style

Streich P, Redwitz J, Walser-Reichenbach S, Herr CEW, Elsner M, Seidel M. Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry. Applied Microbiology. 2024; 4(1):284-296. https://doi.org/10.3390/applmicrobiol4010019

Chicago/Turabian Style

Streich, Philipp, Johannes Redwitz, Sandra Walser-Reichenbach, Caroline E. W. Herr, Martin Elsner, and Michael Seidel. 2024. "Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry" Applied Microbiology 4, no. 1: 284-296. https://doi.org/10.3390/applmicrobiol4010019

APA Style

Streich, P., Redwitz, J., Walser-Reichenbach, S., Herr, C. E. W., Elsner, M., & Seidel, M. (2024). Culture-Independent Quantification of Legionella pneumophila in Evaporative Cooling Systems Using Immunomagnetic Separation Coupled with Flow Cytometry. Applied Microbiology, 4(1), 284-296. https://doi.org/10.3390/applmicrobiol4010019

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