Abstract
World-wide legionellosis outbreaks caused by evaporative cooling systems have shown that there is a need for rapid screening methods for Legionella pneumophila in water. Antibody-based methods for the quantification of L. pneumophila are rapid, non-laborious, and relatively cheap but not sensitive enough for establishment as a screening method for surface and drinking water. Therefore, preconcentration methods have to be applied in advance to reach the needed sensitivity. In a basic test, monolithic adsorption filtration (MAF) was used as primary preconcentration method that adsorbs L. pneumophila with high efficiency. Ten-liter water samples were concentrated in 10 min and further reduced to 1 mL by centrifugal ultrafiltration (CeUF). The quantification of L. pneumophila strains belonging to the monoclonal subtype Bellingham was performed via flow-based chemiluminescence sandwich microarray immunoassays (CL-SMIA) in 36 min. The whole analysis process takes 90 min. A polyclonal antibody (pAb) against L. pneumophila serogroup 1–12 and a monoclonal antibody (mAb) against L. pneumophila SG 1 strain Bellingham were immobilized on a microarray chip. Without preconcentration, the detection limit was 4.0 × 103 and 2.8 × 103 CFU/mL determined by pAb and mAb 10/6, respectively. For samples processed by MAF-CeUF prior to SMIA detection, the limit of detection (LOD) could be decreased to 8.7 CFU/mL and 0.39 CFU/mL, respectively. A recovery of 99.8 ± 15.9 % was achieved for concentrations between 1–1000 CFU/mL. The established combined analytical method is sensitive for rapid screening of surface and drinking water to allow fast hygiene control of L. pneumophila.
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Muder RR, Yu VL, Woo AH. Arch Intern Med. 1986;146:1607–12.
Atlas RM. Environ Microbiol. 1999;61:283–93.
Rangel KM, Delclos G, Emery R, Symanski E. J Occup Environ Hyg. 2011;8:249–65.
Fields BS, Benson RF, Besser RE. Clin Microbiol Rev. 2002;15:506–26.
Fliermans C, Cherry W, Orrison L, Smith S, Tison D, Pope D. Appl Environ Microbiol. 1981;41:9–16.
Brooks T, Osicki RA, Springthorpe VS, Sattar SA, Filion L, Abrial D, et al. J Toxicol Environ Health A. 2004;67:1845–59.
Abdel-Nour M, Duncan C, Low DE, Guyard C. Int J Mol Sci. 2013;14:21660–75.
van Heijnsbergen E, Schalk JAC, Euser SM, Brandsema PS, den Boer JW, Husman AMD. Environ Sci Technol. 2015;49:4797–815.
Helbig JM, Kurtz JB, Pastoris MC, Pelaz C, Lück PC. J Clin Microbiol. 1997;35:2841–5.
Joly JR, McKinney RM, Tobin JO, Bibb WF, Watkins ID, Ramsay D. J Clin Microbiol. 1986;23:76–771.
Kunze A, Pei L, Elsaesser D, Niessner R, Seidel M. J Virol Methods. 2015;222:132–7.
Wolter A, Niessner R, Seidel M. Anal Chem. 2008;80:5854–63.
Kloth K, Niessner R, Seidel M. Biosens Bioelectron. 2009;24:2106–12.
Lengger S, Otto J, Elsaesser D, Schneider O, Tiehm A, Fleischer J, et al. Anal Bioanal Chem. 2014;406:3323–34.
Trinkwasserverordnung. Bundesministerium der Justiz und für Verbraucherschutz, Berlin.2011.
OSHA. OSHA technical manual section III: chapter 7 Legionnaires’ disease. Washington, DC: Occupational Safety and Health Administration of the US Department of Labor; 1999.
Lee JV, Lai S, Exner M, Lenz J, Gaia V, Casati S, et al. J Appl Microbiol. 2011;110:1032–44.
Fuchslin HP, Kotzsch S, Keserue HA, Egli T. Cytometry. 2010;77:264–74.
Keserue HA, Baumgartner A, Felleisen R, Egli T. Microb Biotechnol. 2012;5:753–63.
Shimada T, Noguchi Y, Jackson JL, Miyashita J, Hayashino Y, Kamiya T, et al. Chest J. 2009;136:1576–85.
Dennis PJL, Wright AE, Rutter DA, Death JE, Jones BPC. J Hyg Cambridge. 1984;93:34–353.
Peskoller C, Niessner R, Seidel M. J Chromatogr A. 2009;1216:3794–801.
Karsunke XY, Niessner R, Seidel M. Anal Bioanal Chem. 2009;395:1623–30.
Langer V, Hartmann G, Niessner R, Seidel M. J Aerosol Sci. 2012;48:46–55.
Wutz K, Meyer VK, Wachek S, Krol P, Gareis M, Nölting C, et al. Anal Chem. 2013;85:5279–85.
Blatny JM, Reif BA, Skogan G, Andreassen O, Høiby EA, Ask E, et al. Environ Sci Technol. 2008;42:7360–7.
Leoni E, Legnani P. J Appl Microbiol. 2001;90:27–33.
Wery N, Bru-Adan V, Minervini C, Delgenes JP, Garrelly L, Godon JJ. Appl Environ Microbiol. 2008;74:3030–7.
Kirschner AK, Rameder A, Schrammel B, Indra A, Farnleitner AH, Sommer R. J Appl Microbiol. 2012;112:1244–56.
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Wunderlich, A., Torggler, C., Elsässer, D. et al. Rapid quantification method for Legionella pneumophila in surface water. Anal Bioanal Chem 408, 2203–2213 (2016). https://doi.org/10.1007/s00216-016-9362-x
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DOI: https://doi.org/10.1007/s00216-016-9362-x