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A Rapid and Sensitive CRISPR-Cas12a for the Detection of Legionella pneumophila Cover

A Rapid and Sensitive CRISPR-Cas12a for the Detection of Legionella pneumophila

Open Access
|Dec 2025

References

  1. Abu Khweek A, Amer AO. Factors mediating environmental biofilm formation by Legionella pneumophila. Front Cell Infect Microbiol. 2018 Feb;8:38. https://doi.org/10.3389/fcimb.2018.00038
  2. Ahmed S, Liwak-Muir U, Walker D, Zoldowski A, Mears A, Golovan S, Mohr S, Lem P, Harder C. Validation and in-field testing of a new on-site qPCR system for quantification of Legionella pneumophila according to ISO/TS 12869:2012 in HVAC cooling towers. J Water Health. 2019 Apr;17(2):237-253. https://doi.org/10.2166/wh.2019.252
  3. Avni T, Bieber A, Green H, Steinmetz T, Leibovici L, Paul M. Diagnostic accuracy of PCR alone and compared to urinary antigen testing for detection of Legionella spp.: A systematic review. J Clin Microbiol. 2016 Feb;54(2):401-411. https://doi.org/10.1128/JCM.02675-15
  4. Behets J, Declerck P, Delaedt Y, Creemers B, Ollevier F. Development and evaluation of a Taqman duplex real-time PCR quantification method for reliable enumeration of Legionella pneumophila in water samples. J Microbiol Methods. 2007 Jan;68(1):137-144. https://doi.org/10.1016/j.mimet.2006.07.002
  5. Bonetta S, Bonetta S, Ferretti E, Balocco F, Carraro E. Evaluation of Legionella pneumophila contamination in Italian hotel water systems by quantitative real-time PCR and culture methods. J Appl Microbiol. 2010 May;108(5):1576-1583. https://doi.org/10.1111/j.1365-2672.2009.04553.x
  6. Broughton JP, Deng X, Yu G, Fasching CL, Servellita V, Singh J, Miao X, Streithorst JA, Granados A, Sotomayor-Gonzalez A, et al. CRISPR-Cas12-based detection of SARS-CoV-2. Nat Biotechnol. 2020 Jul;38(7):870-874. https://doi.org/10.1038/s41587-020- 0513-4
  7. Burillo A, Pedro-Botet ML, Bouza E. Microbiology and Epidemiology of Legionnaire’s Disease. Infect Dis Clin North Am. 2017 Mar;31(1):7-27. https://doi.org/10.1016/j.idc.2016.10.002
  8. Burlingame GA, Bartrand TA. Laying the groundwork for a Legionella pneumophila risk management program for public drinking water systems. J Water Health. 2024 Dec;22(12):2385-2397. https://doi.org/10.2166/wh.2024.476
  9. Chen JS, Ma E, Harrington LB, Da Costa M, Tian X, Palefsky JM, Doudna JA. CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity. Science. 2018 Apr;360(6387):436-439. https://doi.org/10.1126/science.aar6245
  10. Collins S, Jorgensen F, Willis C, Walker J. Real-time PCR to supplement gold-standard culture-based detection of Legionella in environmental samples. J Appl Microbiol. 2015 Oct;119(4):1158- 1169 https://doi.org/10.1111/jam.12911
  11. Collins S, Stevenson D, Walker J, Bennett A. Evaluation of Legionella real-time PCR against traditional culture for routine and public health testing of water samples. J Appl Microbiol. 2017 Jun;122(6):1692-1703. https://doi.org/10.1111/jam.13461
  12. Falzone L, Gattuso G, Lombardo C, Lupo G, Grillo CM, Spandidos DA, Libra M, Salmeri M. Droplet digital PCR for the detection and monitoring of Legionella pneumophila. Int J Mol Med. 2020 Nov;46(5):1777-1782. https://doi.org/10.3892/ijmm.2020.4724
  13. Fang Z, Zhou X, Liao H, Xu H. A meta-analysis of Legionella pneumophila contamination in hospital water systems. Am J Infect Control. 2023 Nov;51(11):1250-1262. https://doi.org/10.1016/j. ajic.2023.04.002
  14. Fittipaldi M, Codony F, Morató J. Comparison of conventional culture and real time quantitative PCR using SYBR Green for detection of Legionella pneumophila in water samples. Water SA 2010;36(4):417-24. https://doi.org/10.4314/wsa.v36i4.58411
  15. Gong X, Li J, Zhang Y, Hou S, Qu P, Yang Z, Chen S. Molecular typing of Legionella pneumophila from air-conditioning cooling waters using mip gene, SBT, and FAFLP methods. J Microbiol Methods. 2017 Aug;139:1-7. https://doi.org/10.1016/j.mimet.2017.04.002
  16. Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung J, Verdine V, Donghia N, Daringer NM, Freije CA, et al. Nucleic acid detection with CRISPR-Cas13a/C2c2. Science. 2017 Apr;356(6336):438-442. https://doi.org/10.1126/science.aam9321
  17. Graham CI, MacMartin TL, de Kievit TR, Brassinga AKC. Molecular regulation of virulence in Legionella pneumophila. Mol Microbiol. 2024 Feb;121(2):167-195. https://doi.org/10.1111/mmi.15172
  18. He J, Huang F, Chen H, Chen Q, Zhang J, Li J, Chen D, Chen J. Recombinant Mip-PilE-FlaA dominant epitopes vaccine candidate against Legionella pneumophila. Immunol Lett. 2017 Jun;186:33- 40. https://doi.org/10.1016/j.imlet.2017.03.016
  19. He J, Zhang J, He Y, Huang F, Li J, Chen Q, Chen D, Chen J. Construction of recombinant Mip-FlaA dominant epitope vaccine against Legionella pneumophila and evaluation of the immunogenicity and protective immunity. Immunol Res. 2016 Feb;64(1):272- 279. https://doi.org/10.1007/s12026-015-8746-x
  20. Herwaldt LA, Marra AR. Legionella: A reemerging pathogen. Curr Opin Infect Dis. 2018 Aug;31(4):325-333. https://doi.org/10.1097/QCO.0000000000000468
  21. Karagöz MS, Unal CM, Mayer BE, Müsken M, Borrero-de Acuna JM, Steinert M. Legionella pneumophila PPIase Mip interacts with the bacterial proteins SspB, Lpc2061, and FlaA and promotes flagellation. Infect Immun. 2022 Nov;90(11):e0027622. https://doi.org/10.1128/iai.00276-22
  22. Kozel TR, Burnham-Marusich AR. Point-of-care testing for infectious diseases: Past, present, and future. J Clin Microbiol. 2017 Aug;55(8):2313-2320. https://doi.org/10.1128/JCM.00476-17
  23. Krøjgaard LH, Krogfelt KA, Albrechtsen HJ, Uldum SA. Detection of Legionella by quantitative-polymerase chain reaction (qPCR) for monitoring and risk assessment. BMC Microbiol. 2011 Nov;11:254. https://doi.org/10.1186/1471-2180-11-254
  24. Morio F, Corvec S, Caroff N, Le Gallou F, Drugeon H, Reynaud A. Real-time PCR assay for the detection and quantification of Legionella pneumophila in environmental water samples: Utility for daily practice. Int J Hyg Environ Health. 2008 Jul;211(3-4):403- 411. https://doi.org/10.1016/j.ijheh.2007.06.002
  25. Palusińska-Szysz M, Cendrowska-Pinkosz M. Pathogenicity of the family Legionellaceae. Arch Immunol Ther Exp. 2009 Jul-Aug;57(4):279-290. https://doi.org/10.1007/s00005-009-0035-8
  26. Pascale MR, Mazzotta M, Salaris S, Girolamini L, Grottola A, Simone ML, Cordovana M, Bisognin F, Dal Monte P, Bucci Sabattini MA, et al. Evaluation of MALDI-TOF mass spectrometry in diagnostic and environmental surveillance of Legionella species: A comparison with culture and Mip-gene sequencing technique. Front Microbiol. 2020 Dec 15;11:589369. https://doi.org/10.3389/fmicb.2020.589369
  27. Pierre DM, Baron J, Yu VL, Stout JE. Diagnostic testing for Legionnaires’ disease. Ann Clin Microbiol Antimicrob. 2017 Aug;16(1):59. https://doi.org/10.1186/s12941-017-0229-6
  28. Romanov KA, O’Connor TJ. Legionella pneumophila, a Rosetta stone to understanding bacterial pathogenesis. J Bacteriol. 2024 Dec;206(12):e0032424. https://doi.org/10.1128/jb.00324-24
  29. Samhan FA, Stedtfeld TM, Waseem H, Williams MR, Stedtfeld RD, Hashsham SA. On-filter direct amplification of Legionella pneumophila for rapid assessment of its abundance and viability. Water Res. 2017 Sep;121:162-170. https://doi.org/10.1016/j. watres.2017.05.028
  30. Scheithauer L, Karagöz MS, Mayer BE, Steinert M. Protein sociology of ProA, Mip and other secreted virulence factors at the Legionella pneumophila surface. Front Cell Infect Microbiol. 2023 Mar;13:1140688. https://doi.org/10.3389/fcimb.2023.1140688
  31. Soda EA, Barskey AE, Shah PP, Schrag S, Whitney CG, Arduino MJ, Reddy SC, Kunz JM, Hunter CM, Raphael BH, et al. Vital signs: health care-associated Legionnaires’ disease surveillance data from 20 states and a large metropolitan area - United States, 2015. Am J Transplant. 2017 Aug;17(8):2215-2220. https://doi.org/10.1111/ajt.14407
  32. Tan M, Liao C, Liang L, Yi X, Zhou Z, Wei G. Recent advances in recombinase polymerase amplification: Principle, advantages, disadvantages and applications. Front Cell Infect Microbiol. 2022 Nov;12:1019071. https://doi.org/10.3389/fcimb.2022.1019071
  33. Toplitsch D, Platzer S, Pfeifer B, Hautz J, Mascher F, Kittinger C. Legionella detection in environmental samples as an example for successful implementation of qPCR. Water 2018;10(8):1012. https://doi.org/10.3390/w10081012
  34. Toplitsch D, Platzer S, Zehner R, Maitz S, Mascher F, Kittinger C. Comparison of updated methods for Legionella detection in environmental water samples. Int J Environ Res Public Health. 2021 May;18(10):5436. https://doi.org/10.3390/ijerph18105436
  35. Walker JT, McDermott PJ. Confirming the presence of Legionella pneumophila in your water system: A review of current Legionella testing methods. J AOAC Int. 2021 Aug;104(4):1135-1147. https://doi.org/10.1093/jaoacint/qsab003
  36. Xu T, Cao F, Dai T, Liu T. RPA-CRISPR/Cas12a-mediated isothermal amplification for rapid detection of Phytopythium helicoides. Plant Dis. 2024 Dec;108(12):3463-3472. https://doi.org/10.1094/PDIS-06-24-1300-SR
  37. Yaradou DF, Hallier-Soulier S, Moreau S, Poty F, Hillion Y, Reyrolle M, André J, Festoc G, Delabre K, Vandenesch F, et al. Integrated real-time PCR for detection and monitoring of Legionella pneumophila in water systems. Appl Environ Microbiol. 2007 Mar;73(5):1452-1456. https://doi.org/10.1128/AEM.02399-06
  38. Yin X, Chen YZ, Ye QQ, Liao LJ, Cai ZR, Lin M, Li JN, Zhang GB, Peng XL, Shi WF, et al. Detection performance of PCR for Legionella pneumophila in environmental samples: A systematic review and meta-analysis. Ann Clin Microbiol Antimicrob. 2022 Mar;21(1):12. https://doi.org/10.1186/s12941-022-00503-9
DOI: https://doi.org/10.33073/pjm-2025-041 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
Language: English
Page range: 484 - 493
Submitted on: Aug 1, 2025
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Accepted on: Oct 29, 2025
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Published on: Dec 26, 2025
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year

© 2025 JIANGHAO LI, XINYU WANG, XINLING WANG, HAI QU, LIFEI GAO, ZHONGLING ZHAO, PEI LUO, YEHUAN ZHENG, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.