Advertisement

Analytical and Bioanalytical Chemistry

, Volume 408, Issue 9, pp 2203–2213 | Cite as

Rapid quantification method for Legionella pneumophila in surface water

  • Anika Wunderlich
  • Carmen Torggler
  • Dennis Elsässer
  • Christian Lück
  • Reinhard Niessner
  • Michael SeidelEmail author
Paper in Forefront

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.

Graphical Abstract

Combined process with MAF, CeUF and CL-SMIA for the rapid detection of L. pneumophila in surface water

Keywords

Legionella pneumophila Microarray-based immunoassay Preconcentration of bacteria Monolith Surface water 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

References

  1. 1.
    Muder RR, Yu VL, Woo AH. Arch Intern Med. 1986;146:1607–12.CrossRefGoogle Scholar
  2. 2.
    Atlas RM. Environ Microbiol. 1999;61:283–93.CrossRefGoogle Scholar
  3. 3.
    Rangel KM, Delclos G, Emery R, Symanski E. J Occup Environ Hyg. 2011;8:249–65.CrossRefGoogle Scholar
  4. 4.
    Fields BS, Benson RF, Besser RE. Clin Microbiol Rev. 2002;15:506–26.CrossRefGoogle Scholar
  5. 5.
    Fliermans C, Cherry W, Orrison L, Smith S, Tison D, Pope D. Appl Environ Microbiol. 1981;41:9–16.Google Scholar
  6. 6.
    Brooks T, Osicki RA, Springthorpe VS, Sattar SA, Filion L, Abrial D, et al. J Toxicol Environ Health A. 2004;67:1845–59.CrossRefGoogle Scholar
  7. 7.
    Abdel-Nour M, Duncan C, Low DE, Guyard C. Int J Mol Sci. 2013;14:21660–75.CrossRefGoogle Scholar
  8. 8.
    van Heijnsbergen E, Schalk JAC, Euser SM, Brandsema PS, den Boer JW, Husman AMD. Environ Sci Technol. 2015;49:4797–815.CrossRefGoogle Scholar
  9. 9.
    Helbig JM, Kurtz JB, Pastoris MC, Pelaz C, Lück PC. J Clin Microbiol. 1997;35:2841–5.Google Scholar
  10. 10.
    Joly JR, McKinney RM, Tobin JO, Bibb WF, Watkins ID, Ramsay D. J Clin Microbiol. 1986;23:76–771.Google Scholar
  11. 11.
    Kunze A, Pei L, Elsaesser D, Niessner R, Seidel M. J Virol Methods. 2015;222:132–7.CrossRefGoogle Scholar
  12. 12.
    Wolter A, Niessner R, Seidel M. Anal Chem. 2008;80:5854–63.CrossRefGoogle Scholar
  13. 13.
    Kloth K, Niessner R, Seidel M. Biosens Bioelectron. 2009;24:2106–12.CrossRefGoogle Scholar
  14. 14.
    Lengger S, Otto J, Elsaesser D, Schneider O, Tiehm A, Fleischer J, et al. Anal Bioanal Chem. 2014;406:3323–34.CrossRefGoogle Scholar
  15. 15.
    Trinkwasserverordnung. Bundesministerium der Justiz und für Verbraucherschutz, Berlin.2011.Google Scholar
  16. 16.
    OSHA. OSHA technical manual section III: chapter 7 Legionnaires’ disease. Washington, DC: Occupational Safety and Health Administration of the US Department of Labor; 1999.Google Scholar
  17. 17.
    Lee JV, Lai S, Exner M, Lenz J, Gaia V, Casati S, et al. J Appl Microbiol. 2011;110:1032–44.CrossRefGoogle Scholar
  18. 18.
    Fuchslin HP, Kotzsch S, Keserue HA, Egli T. Cytometry. 2010;77:264–74.Google Scholar
  19. 19.
    Keserue HA, Baumgartner A, Felleisen R, Egli T. Microb Biotechnol. 2012;5:753–63.CrossRefGoogle Scholar
  20. 20.
    Shimada T, Noguchi Y, Jackson JL, Miyashita J, Hayashino Y, Kamiya T, et al. Chest J. 2009;136:1576–85.CrossRefGoogle Scholar
  21. 21.
    Dennis PJL, Wright AE, Rutter DA, Death JE, Jones BPC. J Hyg Cambridge. 1984;93:34–353.CrossRefGoogle Scholar
  22. 22.
    Peskoller C, Niessner R, Seidel M. J Chromatogr A. 2009;1216:3794–801.CrossRefGoogle Scholar
  23. 23.
    Karsunke XY, Niessner R, Seidel M. Anal Bioanal Chem. 2009;395:1623–30.CrossRefGoogle Scholar
  24. 24.
    Langer V, Hartmann G, Niessner R, Seidel M. J Aerosol Sci. 2012;48:46–55.CrossRefGoogle Scholar
  25. 25.
    Wutz K, Meyer VK, Wachek S, Krol P, Gareis M, Nölting C, et al. Anal Chem. 2013;85:5279–85.CrossRefGoogle Scholar
  26. 26.
    Blatny JM, Reif BA, Skogan G, Andreassen O, Høiby EA, Ask E, et al. Environ Sci Technol. 2008;42:7360–7.CrossRefGoogle Scholar
  27. 27.
    Leoni E, Legnani P. J Appl Microbiol. 2001;90:27–33.CrossRefGoogle Scholar
  28. 28.
    Wery N, Bru-Adan V, Minervini C, Delgenes JP, Garrelly L, Godon JJ. Appl Environ Microbiol. 2008;74:3030–7.CrossRefGoogle Scholar
  29. 29.
    Kirschner AK, Rameder A, Schrammel B, Indra A, Farnleitner AH, Sommer R. J Appl Microbiol. 2012;112:1244–56.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Anika Wunderlich
    • 1
  • Carmen Torggler
    • 1
  • Dennis Elsässer
    • 1
  • Christian Lück
    • 2
  • Reinhard Niessner
    • 1
  • Michael Seidel
    • 1
    Email author
  1. 1.Chair of Analytical Chemistry and Institute of HydrochemistryTechnical University of MunichMunichGermany
  2. 2.Institute of Medical Microbiology and Hygiene, Institute of Virology, Medical Faculty “C. G. Carus”Technical University of DresdenDresdenGermany

Personalised recommendations