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Droplet digital PCR for the detection and monitoring of Legionella pneumophila

  • Authors:
    • Luca Falzone
    • Giuseppe Gattuso
    • Cinzia Lombardo
    • Gabriella Lupo
    • Caterina Maria Grillo
    • Demetrios A. Spandidos
    • Massimo Libra
    • Mario Salmeri
  • View Affiliations / Copyright

    Affiliations: Epidemiology Unit, IRCCS Istituto Nazionale Tumori ‘Fondazione G. Pascale’, I‑80131 Naples, Italy, Department of Biomedical and Biotechnological Sciences, Section of General Pathology, University of Catania, I‑95123 Catania, Italy, Department of Biomedical and Biotechnological Sciences, Section of Microbiology, University of Catania, I‑95123 Catania, Italy, Department of Biomedical and Biotechnological Sciences, Section of Biochemistry, University of Catania, I‑95123 Catania, Italy, Department of Medical Sciences, Surgical and Advanced Technologies, GF Ingrassia, University of Catania, I‑95123 Catania, Italy, Laboratory of Clinical Virology, Medical School, University of Crete, 71003 Heraklion, Greece
    Copyright: © Falzone et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 1777-1782
    |
    Published online on: September 14, 2020
       https://doi.org/10.3892/ijmm.2020.4724
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Abstract

Legionella pneumophila (L. pneumophila) is a harmful pathogen often found in water systems. In hospitals, the absence of L. pneumophila in water systems is mandatory by law, therefore, frequent and effective monitoring of water is of fundamental importance. Molecular methods based on reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) have been proposed for the detection of L. pneumophila, however, the sensitivity and accuracy of these methods have not been validated yet. Therefore, it is important to evaluate other strategies able to overcome the limits of culture‑based and RT‑qPCR methods. On these bases, we compared the sensitivity and accuracy of droplet digital PCR (ddPCR) and RT‑qPCR in water samples with known concentrations of L. pneumophila and in an in vitro model of water heat treatments. ddPCR showed a higher sensitivity rate and accuracy compared to RT‑qPCR in detecting low bacterial load. In addition, ddPCR is not affected by the presence of fragmented DNA and showed higher accuracy than RT‑qPCR in monitoring the efficacy of heat shock treatments. In conclusion, ddPCR represents an innovative strategy to effectively detect L. pneumophila in water samples. Thanks to its high robustness, ddPCR could be applied also for the detection of L. pneumophila in patients with suspected legionellosis.
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1 

Cunha BA, Burillo A and Bouza E: Legionnaires' disease. Lancet. 387:376–385. 2016. View Article : Google Scholar

2 

Stout JE and Yu VL: Legionellosis. N Engl J Med. 337:682–687. 1997. View Article : Google Scholar : PubMed/NCBI

3 

European Centre for Disease Prevention and Control: Legionnaires' Disease in Europe, Annual Epidemiological Report for 2017. ECDC; Stockholm: 2019

4 

Istituto Superiore di Sanità (ISS): Rapporto Annuale sulla Legionellosi in Italia Nel 2017. 31. ISS; Rome: 2018

5 

Bartram J, Chartier Y, Lee JV, Pond K and Surman-Lee S: Legionella and the prevention of legionellosis. World Health Organization Press; Geneva: 2007

6 

Fliermans CB, Soracco RJ and Pope DH: Measure of Legionella pneumophila activity in situ. Curr Microbiol. 6:89–94. 1981. View Article : Google Scholar

7 

Abdel-Nour M, Duncan C, Low DE and Guyard C: Biofilms: The stronghold of Legionella pneumophila. Int J Mol Sci. 14:21660–21675. 2013. View Article : Google Scholar : PubMed/NCBI

8 

Liu Z, Lin YE, Stout JE, Hwang CC, Vidic RD and Yu VL: Effect of flow regimes on the presence of Legionella within the biofilm of a model plumbing system. J Appl Microbiol. 101:437–442. 2006. View Article : Google Scholar : PubMed/NCBI

9 

van Heijnsbergen E, Schalk JA, Euser SM, Brandsema PS, den Boer JW and de Roda Husman AM: Confirmed and potential sources of Legionella reviewed. Environ Sci Technol. 49:4797–4815. 2015. View Article : Google Scholar : PubMed/NCBI

10 

Linee guida per la prevenzione ed il controllo della legionellosi: Conferenza Stato-Regioni nella seduta del 7 maggio 2015. http://www.salute.gov.it/portale/documentazione/p6_2_2_1.jsp?lingua=italiano&id=2362uri. Accessed May 13, 2015.

11 

ISO 11731:2017: Water Quality-Enumeration of Legionella. International Organization for Standardization; Geneva: 2017, https://www.iso.org/standard/61782.htmluri.

12 

Kimura S, Tateda K, Ishii Y, Horikawa M, Miyairi S, Gotoh N, Ishiguro M and Yamaguchi K: Pseudomonas aeruginosa Las quorum sensing autoinducer suppresses growth and biofilm production in Legionella species. Microbiol Read. 155:1934–1939. 2009. View Article : Google Scholar

13 

Leoni E and Legnani PP: Comparison of selective procedures for isolation and enumeration of Legionella species from hot water systems. J Appl Microbiol. 90:27–33. 2001. View Article : Google Scholar : PubMed/NCBI

14 

Ta AC, Stout JE, Yu VL and Wagener MM: Comparison of culture methods for monitoring Legionella species in hospital potable water systems and recommendations for standardization of such methods. J Clin Microbiol. 33:2118–2123. 1995. View Article : Google Scholar : PubMed/NCBI

15 

Touron-Bodilis A, Pougnard C, Frenkiel-Lebossé H and Hallier-Soulier S: Usefulness of real-time PCR as a complementary tool to the monitoring of Legionella spp. and Legionella pneumophila by culture in industrial cooling systems. J Appl Microbiol. 111:499–510. 2011. View Article : Google Scholar : PubMed/NCBI

16 

Bonetta S, Bonetta S, Ferretti E, Balocco F and Carraro E: Evaluation of Legionella pneumophila contamination in Italian hotel water systems by quantitative real-time PCR and culture methods. J Appl Microbiol. 108:1576–1583. 2010. View Article : Google Scholar

17 

Edagawa A, Kimura A, Doi H, Tanaka H, Tomioka K, Sakabe K, Nakajima C and Suzuki Y: Detection of culturable and nonculturable Legionella species from hot water systems of public buildings in Japan. J Appl Microbiol. 105:2104–2114. 2008. View Article : Google Scholar

18 

Lee S and Lee J: Outbreak investigations and identification of Legionella in contaminated water. Methods Mol Biol. 954:87–118. 2013. View Article : Google Scholar

19 

Arvia R, Sollai M, Pierucci F, Urso C, Massi D and Zakrzewska K: Droplet digital PCR (ddPCR) vs quantitative real-time PCR (qPCR) approach for detection and quantification of Merkel cell polyomavirus (MCPyV) DNA in formalin fixed paraffin embedded (FFPE) cutaneous biopsies. J Virol Methods. 246:15–20. 2017. View Article : Google Scholar : PubMed/NCBI

20 

Hayden RT, Gu Z, Ingersoll J, Abdul-Ali D, Shi L, Pounds S and Caliendo AM: Comparison of droplet digital PCR to real-time PCR for quantitative detection of cytomegalovirus. J Clin Microbiol. 51:540–546. 2013. View Article : Google Scholar :

21 

Filetti V, Falzone L, Rapisarda V, Caltabiano R, Eleonora Graziano AC, Ledda C and Loreto C: Modulation of microRNA expression levels after naturally occurring asbestiform fibers exposure as a diagnostic biomarker of mesothelial neoplastic transformation. Ecotoxicol Environ Saf. 198:1106402020. View Article : Google Scholar : PubMed/NCBI

22 

Battaglia R, Palini S, Vento ME, La Ferlita A, Lo Faro MJ, Caroppo E, Borzì P, Falzone L, Barbagallo D, Ragusa M, et al: Identification of extracellular vesicles and characterization of miRNA expression profiles in human blastocoel fluid. Sci Rep. 9:842019. View Article : Google Scholar : PubMed/NCBI

23 

Salemi R, Falzone L, Madonna G, Polesel J, Cinà D, Mallardo D, Ascierto PA, Libra M and Candido S: MMP-9 as a candidate marker of response to BRAF inhibitors in melanoma patients with BRAFV600E mutation detected in circulating-free DNA. Front Pharmacol. 9:8562018. View Article : Google Scholar :

24 

Falzone L, Musso N, Gattuso G, Bongiorno D, Palermo CI, Scalia G, Libra M and Stefani S: Droplet digital PCR as the best sensitive assay for the SARS-CoV-2 detection. Int J Mol Med. 46:957–964. 2020. View Article : Google Scholar : PubMed/NCBI

25 

Krøjgaard LH, Krogfelt KA, Albrechtsen HJ and Uldum SA: Detection of Legionella by quantitative-polymerase chain reaction (qPCR) for monitoring and risk assessment. BMC Microbiol. 11:2542011. View Article : Google Scholar : PubMed/NCBI

26 

Shen SM, Chou MY, Hsu BM, Ji WT, Hsu TK, Tsai HF, Huang YL, Chiu YC, Kao ES, Kao PM, et al: Assessment of Legionella pneumophila in recreational spring water with quantitative PCR (Taqman) assay. Pathog Glob Health. 109:236–241. 2015. View Article : Google Scholar : PubMed/NCBI

27 

Ziegler I, Lindström S, Källgren M, Strålin K and Mölling P: 16S rDNA droplet digital PCR for monitoring bacterial DNAemia in bloodstream infections. PLoS One. 14:e02246562019. View Article : Google Scholar : PubMed/NCBI

28 

Singh G, Sithebe A, Enitan AM, Kumari S, Bux F and Stenström TA: Comparison of droplet digital PCR and quantitative PCR for the detection of Salmonella and its application for river sediments. J Water Health. 15:505–508. 2017. View Article : Google Scholar : PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Falzone L, Gattuso G, Lombardo C, Lupo G, Grillo CM, Spandidos DA, Libra M and Salmeri M: Droplet digital PCR for the detection and monitoring of Legionella pneumophila. Int J Mol Med 46: 1777-1782, 2020.
APA
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C.M., Spandidos, D.A. ... Salmeri, M. (2020). Droplet digital PCR for the detection and monitoring of Legionella pneumophila. International Journal of Molecular Medicine, 46, 1777-1782. https://doi.org/10.3892/ijmm.2020.4724
MLA
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C. M., Spandidos, D. A., Libra, M., Salmeri, M."Droplet digital PCR for the detection and monitoring of Legionella pneumophila". International Journal of Molecular Medicine 46.5 (2020): 1777-1782.
Chicago
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C. M., Spandidos, D. A., Libra, M., Salmeri, M."Droplet digital PCR for the detection and monitoring of Legionella pneumophila". International Journal of Molecular Medicine 46, no. 5 (2020): 1777-1782. https://doi.org/10.3892/ijmm.2020.4724
Copy and paste a formatted citation
x
Spandidos Publications style
Falzone L, Gattuso G, Lombardo C, Lupo G, Grillo CM, Spandidos DA, Libra M and Salmeri M: Droplet digital PCR for the detection and monitoring of Legionella pneumophila. Int J Mol Med 46: 1777-1782, 2020.
APA
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C.M., Spandidos, D.A. ... Salmeri, M. (2020). Droplet digital PCR for the detection and monitoring of Legionella pneumophila. International Journal of Molecular Medicine, 46, 1777-1782. https://doi.org/10.3892/ijmm.2020.4724
MLA
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C. M., Spandidos, D. A., Libra, M., Salmeri, M."Droplet digital PCR for the detection and monitoring of Legionella pneumophila". International Journal of Molecular Medicine 46.5 (2020): 1777-1782.
Chicago
Falzone, L., Gattuso, G., Lombardo, C., Lupo, G., Grillo, C. M., Spandidos, D. A., Libra, M., Salmeri, M."Droplet digital PCR for the detection and monitoring of Legionella pneumophila". International Journal of Molecular Medicine 46, no. 5 (2020): 1777-1782. https://doi.org/10.3892/ijmm.2020.4724
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