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Rapid and specific SPRi detection of L. pneumophila in complex environmental water samples

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Abstract

Legionellosis is a very devastating disease worldwide mainly due to unpredictable outbreaks in man-made water systems. Developing a highly specific and sensitive rapid detection system that detects only metabolically active bacteria is a main priority for water quality assessment. We previously developed a versatile technique for sensitive and specific detection of synthetic RNA. In the present work, we further investigated the performance of the developed biosensor for detection of Legionella pneumophila in complex environmental samples, particularly those containing protozoa. The specificity and sensitivity of the detection system were verified using total RNA extracted from L. pneumophila in spiked water co-cultured with amoebae. We demonstrated that the expression level of ribosomal RNA (rRNA) is extremely dependent on the environmental conditions. The presence of amoebae with L. pneumophila, especially in nutrition-deprived samples, increased the amount of L. pneumophila 15-fold after 1 week as measured through the expression of 16s rRNA. Using the developed surface plasmon resonance imaging (SPRi) detection method, we were also able to successfully detect L. pneumophila within 3 h, both in the presence and absence of amoebae in the complex environmental samples obtained from a cooling water tower. These findings suggest that the developed biosensing system is a viable method for rapid, real-time and effective detection not only for L. pneumophila in environmental samples but also to assess the risk associated with the use of water contaminated with other pathogens.

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References

  1. Swanson M, Hammer B (2000) Legionella pneumophila pathogenesis: a fateful journey from amoebae to macrophages. Ann Rev Microbiol 54(1):567–613

    Article  CAS  Google Scholar 

  2. Craun GF, Brunkard JM, Yoder JS, Roberts VA, Carpenter J, Wade T, Calderon RL, Roberts JM, Beach MJ, Roy SL (2010) Causes of outbreaks associated with drinking water in the United States from 1971 to 2006. Clin Microbiol Rev 23(3):507–528

    Article  CAS  Google Scholar 

  3. Wadowsky R, Wilson T, Kapp N, West A, Kuchta J, Dowling J, Yee R (1991) Multiplication of Legionella spp. in tap water containing Hartmannella vermiformis. Appl Environ Microbiol 57(7):1950–1955

    CAS  Google Scholar 

  4. Lazcka O, Campo FJD, Muñoz FX (2007) Pathogen detection: a perspective of traditional methods and biosensors. Biosens Bioelectron 22(7):1205–1217

    Article  CAS  Google Scholar 

  5. Foudeh AM, Fatanat Didar T, Veres T, Tabrizian M (2012) Microfluidic designs and techniques using lab-on-a-chip devices for pathogen detection for point-of-care diagnostics. Lab Chip 12(18):3249–3266

    Article  CAS  Google Scholar 

  6. Deisingh AK, Thompson M (2002) Detection of infectious and toxigenic bacteria. Analyst 127(5):567–581

    Article  CAS  Google Scholar 

  7. Stinear T, Matusan A, Hines K, Sandery M (1996) Detection of a single viable Cryptosporidium parvum oocyst in environmental water concentrates by reverse transcription-PCR. Appl Environ Microbiol 62(9):3385–3390

    CAS  Google Scholar 

  8. Foudeh AM, Daoud JT, Faucher SP, Veres T, Tabrizian M (2014) Sub-femtomole detection of 16s rRNA from Legionella pneumophila using surface plasmon resonance imaging. Biosens Bioelectron 52:129–135

    Article  CAS  Google Scholar 

  9. Borella P, Guerrieri E, Marchesi I, Bondi M, Messi P (2005) Water ecology of Legionella and protozoan: environmental and public health perspectives. Biotechnol Annu Rev 11:355–380

  10. Barbaree JM, Fields BS, Feeley JC, Gorman GW, Martin WT (1986) Isolation of protozoa from water associated with a legionellosis outbreak and demonstration of intracellular multiplication of Legionella pneumophila. Appl Environ Microbiol 51(2):422–424

    CAS  Google Scholar 

  11. Dupuy M, Mazoua S, Berne F, Bodet C, Garrec N, Herbelin P, Ménard-Szczebara F, Oberti S, Rodier M-H, Soreau S, Wallet F, Héchard Y (2011) Efficiency of water disinfectants against Legionella pneumophila and Acanthamoeba. Water Res 45(3):1087–1094

    Article  CAS  Google Scholar 

  12. Bouyer S, Imbert C, Rodier M-H, Héchard Y (2007) Long-term survival of Legionella pneumophila associated with Acanthamoeba castellanii vesicles. Environ Microbiol 9(5):1341–1344

    Article  CAS  Google Scholar 

  13. Cirillo JD, Falkow S, Tompkins LS (1994) Growth of Legionella pneumophila in Acanthamoeba castellanii enhances invasion. Infect Immun 62(8):3254–3261

    CAS  Google Scholar 

  14. Molmeret M, Horn M, Wagner M, Santic M, Abu Kwaik Y (2005) Amoebae as training grounds for intracellular bacterial pathogens. Appl Environ Microbiol 71(1):20–28

    Article  CAS  Google Scholar 

  15. Gourse RL, Gaal T, Bartlett MS, Appleman JA, Ross W (1996) rRNA transcription and growth rate-dependent regulation of ribosome synthesis in Escherichia coli. Annu Rev Microbiol 50(1):645–677

    Article  CAS  Google Scholar 

  16. Neumeister B, Reiff G, Faigle M, Dietz K, Northoff H, Lang F (2000) Influence of Acanthamoeba castellanii on intracellular growth of different Legionella species in human monocytes. Appl Environ Microbiol 66(3):914–919

    Article  CAS  Google Scholar 

  17. Ohno A, Kato N, Sakamoto R, Kimura S, Yamaguchi K (2008) Temperature-dependent parasitic relationship between Legionella pneumophila and a free-living amoeba (Acanthamoeba castellanii). Appl Environ Microbiol 74(14):4585–4588

    Article  CAS  Google Scholar 

  18. Declerck P, Behets J, Delaedt Y, Margineanu A, Lammertyn E, Ollevier F (2005) Impact of non-Legionella bacteria on the uptake and intracellular replication of Legionella pneumophila in Acanthamoeba castellanii and Naegleria lovaniensis. Microb Ecol 50(4):536–549

    Article  CAS  Google Scholar 

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Acknowledgments

We acknowledge the National Science and Engineering Research Council of Canada-Strategic and Discovery programme, Genome Canada/Genome Quebec and Nano-Quebec. The authors would also like to thank R. Tien Sing Young for helping with illustrations design, L. Li for the amoeba culture, Dr. K. Bowey for her comments on the manuscript and Dr. Sandra Imbeault for providing the environmental water samples. The work in the SPF laboratory is supported by NSERC Discovery Grant 418289-2012.

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Correspondence to Maryam Tabrizian.

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Foudeh, A.M., Trigui, H., Mendis, N. et al. Rapid and specific SPRi detection of L. pneumophila in complex environmental water samples. Anal Bioanal Chem 407, 5541–5545 (2015). https://doi.org/10.1007/s00216-015-8726-y

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  • DOI: https://doi.org/10.1007/s00216-015-8726-y

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