Typing of Legionella pneumophila and its Role in Elucidating the Epidemiology of Legionnaires’ Disease

Timothy G. Harrison; Norman K. Fry; Baharak Afshar; William Bellamy; Nita Doshi; Anthony P. Underwood

doi:10.1128/9781555815660.ch25

Chapter 25 : Typing of Legionella pneumophila and its Role in Elucidating the Epidemiology of Legionnaires’ Disease

Authors: Timothy G. Harrison¹, Norman K. Fry², Baharak Afshar³, William Bellamy⁴, Nita Doshi⁵, Anthony P. Underwood⁶

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Affiliations: 1: Health Protection Agency, Respiratory and Systemic Infection Laboratory, Center for Infections, 61 Colindale Ave., London NW9 5EQ; 2: Health Protection Agency, Respiratory and Systemic Infection Laboratory, Center for Infections, 61 Colindale Ave., London NW9 5EQ; 3: Health Protection Agency, Respiratory and Systemic Infection Laboratory, Center for Infections, 61 Colindale Ave., London NW9 5EQ; 4: Health Protection Agency, Respiratory and Systemic Infection Laboratory, Center for Infections, 61 Colindale Ave., London NW9 5EQ; 5: Health Protection Agency, Respiratory and Systemic Infection Laboratory, Center for Infections, 61 Colindale Ave., London NW9 5EQ; 6: Health Protection Agency, Statistics, Modelling and Bioinformatics Department, Center for Infections, 61 Colindale Ave., London NW9 5EQ

Content Type: Monograph

Publication Year: 2006

Category: Bacterial Pathogenesis

Book DOI: 10.1128/9781555815660

Chapter DOI: 10.1128/9781555815660.ch25

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Abstract:

The main reason for typing Legionella pneumophila is to help identify environmental sources giving rise to cases of legionellosis, thus allowing control measures to be implemented and further cases to be prevented in a timely manner. Monoclonal antibody (mAB) subgrouping is probably the most rapid and reliable phenotypic method available and is excellent for initial screening of isolates. Only a few of the many genotypic methods described have been shown to meet these exacting requirements. Of these, pulsed-field gel electrophoresis is the most widely applied, but others have also been used. For example, a restriction-fragment length polymorphism (RFLP) typing method was used as the standard method in the United Kingdom for almost 20 years. Investigation of travel-associated legionellosis requires even more rigorously standardized systems, as not only must the types be clearly defined but they also must be recognizable and reproducible in laboratories in different countries (different laboratories, different times). The standardized sequence-based typing (SBT) method targets part of six genes (flaA, pilE, asd, mip, mompS, proA), each of which shows considerable allelic variation, thus providing an extremely discriminatory typing system. Furthermore, of the seven major-outbreak-associated strains for which SBT data are available, five are SBT 3,4,1,1,14,9. However, if substantiated, these observations have important and widespread implications for our understanding of the epidemiology of legionellosis and for the monitoring and control of L. pneumophila in the built environment.

Citation: G. Harrison T, K. Fry N, Afshar B, Bellamy W, Doshi N, P. Underwood A. 2006. Typing of Legionella pneumophila and its Role in Elucidating the Epidemiology of Legionnaires’ Disease, p 94-99. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), Legionella. ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch25

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Typing of Legionella pneumophila and its Role in Elucidating the Epidemiology of Legionnaires’ Disease

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/content/10.1128/9781555815660.ch25.ch25fig01

FIGURE 1

Minimum spanning tree of 124 unrelated L. pneumophila isolates from the EWGLI culture collection.

10.1128/9781555815660/f0096-01_thmb.gif

10.1128/9781555815660/f0096-01.gif

FIGURE 1

Minimum spanning tree of 124 unrelated L. pneumophila isolates from the EWGLI culture collection.

References

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1. Aurell, H.,, J. Etienne,, F. Forey,, M. Reyrolle,, P. Girardo,, P. Farge,, B. Decludt,, C. Campese,, F. Vandenesch, and, S. Jarraud. 2003. Legionella pneumophila serogroup 1 strain Paris: endemic distribution throughout France. J. Clin. Microbiol. 41: 3320– 3322.

2. Aurell, H.,, P. Farge,, H. Meugnier,, M. Gouy,, F. Forey,, G. Lina,, F. Vandenesch,, J. Etienne, and, S. Jarraud. 2005. Clinical and environmental isolates of Legionella pneumophila serogroup 1 cannot be distinguished by sequence analysis of two surface protein genes and three housekeeping genes. Appl. Environ. Microbiol. 71: 282– 289.

3. Bangsborg, J. M.,, P. Gerner-Smidt,, H. Colding,, N.-E. Fiehn,, B. Bruun, and, N. Høiby. 1995. Restriction fragment length polymorphism of rRNA genes for molecular typing of members of the family Legionellaceae. J. Clin. Microbiol. 33: 402– 406.

4. Bernander, S.,, K. Jacobson,, J. H. Helbig,, P. C. Lück, and, M. Lundholm. 2003. A hospital-associated outbreak of Legionnaires’ disease caused by Legionella pneumophila serogroup 1 is characterized by stable genetic fingerprinting but variable monoclonal antibody patterns. J. Clin. Microbiol. 41: 2503– 2508.

5. Brown, C. M.,, P. J. Nuorti,, R. F. Breiman,, A. L. Hathcock,, B. S. Fields,, H. B. Lipman,, G. C. Llewellyn,, J. Hofmann, and, M. Cetron. 1999. A community outbreak of Legionnaires’ disease linked to hospital cooling towers: an epidemiological method to calculate dose of exposure. Int. J. Epidemiol. 28: 353– 359.

6. Fry, N. K.,, S. Alexiou-Daniel,, J. M. Bangsborg,, S. Bernander,, M. Castellani-Pastoris,, J. Etienne,, B. Forsblom,, V. Gaia,, J. H. Helbig,, D. Lindsay,, L. P. Christian,, P. C. Lück,, C. Pelaz,, S. A. Uldum, and, T. G. Harrison. 1999. A multicenter evaluation of genotypic methods for the epidemiologic typing of Legionella pneumophila serogroup 1: results of a pan-European study. Clin. Microbiol. Infect. 5: 462– 477.

7. Fry, N. K.,, J. M. Bangsborg,, A. Bergmans,, S. Bernander,, J. Etienne,, L. Franzin,, V. Gaia,, P. Hasenberger,, B. Baladrón Jiménez,, D. Jonas,, D. Lindsay,, S. Mentula,, A. Papoutsi,, M. Struelens,, S. A. Uldum,, P. Visca,, W. Wannet, and, T. G. Harrison. 2002. Designation of the European Working Group on Legionella Infection (EWGLI) amplified fragment length polymorphism types of Legionella pneumophila serogroup 1 and results of intercentre proficiency testing using a standard protocol. Eur. J. Clin. Microbiol. Infect. Dis. 21: 722– 728.

8. Gaia, V.,, N. K. Fry,, T. G. Harrison, and, R. Peduzzi. 2003. Sequence-based typing of Legionella pneumophila serogroup 1 offers the potential for true portability in legionellosis outbreak investigation. J. Clin. Microbiol. 41: 2932– 2939.

9. Gaia, V.,, N. K. Fry,, B. Afshar,, P. C. Lück,, H. Meugnier,, J. Etienne,, R. Peduzzi, and, T. G. Harrison. 2005. Consensus sequence-based scheme for epidemiological typing of clinical and environmental isolates of Legionella pneumophila. J. Clin. Microbiol. 43: 2047– 2052.

10. Harrison, T. G.,, N. A. Saunders,, A. Haththotuwa,, N. Doshi, and, A. G. Taylor. 1990. Typing of Legionella pneumophila serogroups 2-14 strains by analysis of restriction fragment length polymorphisms. Lett.Appl. Microbiol. 11: 189– 192.

11. Harrison, T. G.,, N. Doshi,, N. K. Fry, and, C.A. Joseph. 2002. Subtyping of Legionella pneumophila: a review of 19 years of data from England and Wales. Proceedings of the 22nd Annual Meeting of the European Working Group on Legionella Infections. 26– 28 May 2002, St Julian’s, Malta, p. 28.

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13. Joseph, C. A. 2004. Legionnaires’ disease in Europe 2000-2002. Epidemiol. Infect. 132: 417– 424.

14. Lawrence, C.,, M. Reyrolle,, S. Dubrou,, F. Forey,, B. Decludt,, C. Goulvestre,, P. Matsiota-Bernard,, J. Etienne, and, C. Nauciel. 1999. Single clonal origin of a high proportion of Legionella pneumophila serogroup 1 isolates from patients and the environment in the area of Paris, France, over a 10-year period. J. Clin. Microbiol. 37: 2652– 2655.

15. Lück, P. C.,, J. H. Helbig,, H.-J. Hagedorn, and, W. Ehret. 1995. DNA fingerprinting by pulsed-field gel electrophoresis to investigate a nosocomial pneumonia caused by Legionella bozemanii serogroup 1. Appl. Environ. Microbiol. 61: 2759– 2761.

16. Rangel-Frausto, M. S.,, P. Rhomberg,, R. J. Hollis,, M. A. Pfaller,, R. P. Wenzel,, C. M. Helms, and, L. A. Herwaldt. 1999. Persistence of Legionella pneumophila in a hospital’s water system: a 13-year survey. Infect. Control. Hosp. Epidemiol. 20: 793– 797.

17. Selander, R. K.,, R. M. McKinney,, T. S. Whittam,, W. F. Bibb,, D. J. Brenner,, F. S. Nolte, and, P. E. Pattison. 1985. Genetic structure of populations of Legionella pneumophila. J. Bacteriol. 163:1021–1037.

18. van Ketel, R. J.,, J. ter Schegget, and, H. C. Zanen. 1984. Molecular epidemiology of Legionella pneumophila serogroup 1. J. Clin. Microbiol. 20: 362– 364.

Tables

Typing of Legionella pneumophila and its Role in Elucidating the Epidemiology of Legionnaires’ Disease

/content/10.1128/9781555815660.ch25.ch25tab01

TABLE 1

Examples of global distribution of some L. pneumophila clones

TABLE 1

Examples of global distribution of some L. pneumophila clones

/content/10.1128/9781555815660.ch25.ch25tab02

TABLE 2

Available SBT data for L. pneumophila strains causing major outbreaks

TABLE 2

Available SBT data for L. pneumophila strains causing major outbreaks

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