Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years

Robert F. Benson; Claressa E. Lucas; Ellen W. Brown; Karen D. Cowgill; Barry S. Fields

doi:10.1128/9781555815660.ch37

Chapter 37 : Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years

Authors: Robert F. Benson¹, Claressa E. Lucas², Ellen W. Brown³, Karen D. Cowgill⁴, Barry S. Fields⁵

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Affiliations: 1: Respiratory Disease Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, GA 30333; 2: Respiratory Disease Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, GA 30333; 3: Respiratory Disease Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, GA 30333; 4: Seattle Biomedical Research Institute, 307 Westlake Ave N, Suite 500, Seattle WA 98109; 5: Respiratory Disease Branch, Centers for Disease Control and Prevention, 1600 Clifton Rd., Atlanta, GA 30333

Content Type: Monograph

Publication Year: 2006

Category: Bacterial Pathogenesis

Book DOI: 10.1128/9781555815660

Chapter DOI: 10.1128/9781555815660.ch37

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Abstract:

During outbreak investigations molecular analytical methods are necessary to discriminate among clinical isolates and to establish a link with environmental isolates. In addition to monoclonal antibodies used as an initial subtyping method, other methods employed have been pulse-field gel electrophoresis, amplified fragment length polymorphosim (AFLP), arbitrarily primed PCR, multilocus variable-number tandem repeat, and sequence-based typing (SBT). In the present study, the authors used SBT to study the relationships among clinical and environmental isolates of Legionella pneumophila isolated from the same hotel during a 22-year period. Water samples were collected from the hotel and cultured using standard procedures for the recovery of Legionella isolates from environmental sources. To determine the discriminatory power of the SBT procedure, eight isolates that share the same monoclonal antibody (MAb) subtype were included. These isolates represent four enzyme types based on previous studies using multilocus enzyme electrophoresis (MLEE). All isolates were tested with a panel of MAbs to determine their subtype. Isolates were compared by AFLP. Patient isolates and representative environmental isolates from both outbreaks were analyzed by a sequence-based typing scheme.

Citation: F. Benson R, E. Lucas C, W. Brown E, D. Cowgill K, S. Fields B. 2006. Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years, p 139-142. 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.ch37

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Multilocus Sequence Typing: 0.52651095
Legionella pneumophila: 0.5236079

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Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years

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/content/10.1128/9781555815660.ch37.ch37fig01

FIGURE 1

Genetic relatedness of St. Croix patient isolates from 1981 and 2003 and reference Legionella pneumophila serogroup 1 MAb 1,2,3 pattern.

10.1128/9781555815660/f0140-01_thmb.gif

10.1128/9781555815660/f0140-01.gif

FIGURE 1

Genetic relatedness of St. Croix patient isolates from 1981 and 2003 and reference Legionella pneumophila serogroup 1 MAb 1,2,3 pattern.

References

/content/book/10.1128/9781555815660.ch37

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. Foey,, 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. Benson, R. F.,, B. S. Fields,, A. Benin,, H. Craddock, and, R. E. Besser. 2002. Application of amplified fragment length polymorphism analysis to subtyping of Legionella pneumophila serogroup 6, p. 243– 247. In R. Marre,, Y. Abu Kwaik,, C. Bartlett,, N. P. Cianciotto,, B. S. Fields,, M. Frosch,, J. Hacker,, P. C. Luck (ed.). Legionella. ASM Press, Washington, D.C.

4. Darelid, J.,, S. Bernander,, K. Jacobson, and, S. Lofgren. 2004. The presence of a specific genotype of Legionella pneumophila serogroup 1 in a hospital and municipal water distribution system over a 12-year period. Scand. J. Infect. Dis. 36: 417– 423.

5. Gaia, V.,, N. K. Fry,, B. Afshar,, P. C. Luck, et al. 2005. Consensus sequence-based scheme for epidemiological typing of clinical and environmental isolates of Legionella pneumophila. J. Clin. Microbiol. 43: 2047– 2052.

6. Kumar, S.,, K. Tamura, and, M. Nei. 2004. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment. Briefings Bioinformatics 5: 150– 163.

7. Pruckler, J. M.,, L. A. Mermel,, R. F. Benson,, C. Giorgio,, P. K. Cassiday,, R. F. Breiman,, C. G. Whitney, and, B. S. Fields. 1995. Comparison of Legionella pneumophila isolates by arbitrarily primed PCR and pulsed-field gel electrophoresis: analysis from seven epidemic investigations. J. Clin. Microbiol. 33: 2872– 2875.

8. 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.

9. 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.

Tables

Molecular Comparison of Isolates from a Recurring Outbreak of Legionnaires’ Disease Spanning 22 Years

/content/10.1128/9781555815660.ch37.ch37tab01

TABLE 1

SBT for isolates obtained from St. Croix, U.S.Virgin Islands during 1981 and 2003 epidemic investigations and reference strains

TABLE 1

SBT for isolates obtained from St. Croix, U.S.Virgin Islands during 1981 and 2003 epidemic investigations and reference strains

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