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Chapter 18 : Serotyping of in Epidemiological Investigations: Limitations in the Era of Genotyping

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Serotyping of in Epidemiological Investigations: Limitations in the Era of Genotyping, Page 1 of 2

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Abstract:

The end of the 1980s was the “golden age” of serotyping. In recent years molecular genotyping methods have been increasingly applied to the classification of legionellae. This chapter describes a short typing system for that might be useful in epidemiological investigations. Serotyping of is mostly done by using indirect immunofluorescence tests. However, in a European multicenter study for typing of , it was demonstrated that this technique produces variable results, especially for the recognition of serogroup-cross-reactive epitopes by reagents LpH and LpK. These problems can be avoided by using an enzyme-linked immunosorbent assay (ELISA) that is a more reproducible technique because of reading of positive results with respect to the blank value, whereas microscopically judging is sometimes equivocal and influenced by the laboratory staff. ELISA allows the screening of many colonies in less than six hours. In addition, the selection of only 11 reagents combined with a blank without monoclonal antibodies (MAbs) as the 12th value was done to adapt the typing scheme to the microplate array.

Citation: Helbig J, Lück P. 2006. Serotyping of in Epidemiological Investigations: Limitations in the Era of Genotyping, p 68-72. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch18

Key Concept Ranking

Enzyme-Linked Immunosorbent Assay
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Legionella pneumophila
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Figures

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FIGURE 1

Distribution of serogroups and MAb subgroups among 1,335 human isolates (Reprinted from Ref. 5 with kind permission of Springer Science and Business Media). Abbreviations for monoclonal subgroups of serogroup 1: Ph, Philadelphia; Be, Benidorm; FA, France/Allentown; Kn, Knoxville; OO, OLDA/Oxford; He, Heysham; Ca, Camoerdown; Bh, Bellingham. n.t., not typeable into the known serogroups.

Citation: Helbig J, Lück P. 2006. Serotyping of in Epidemiological Investigations: Limitations in the Era of Genotyping, p 68-72. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch18
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References

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1. Brenner, T.U.,, A. G. Steigerwalt,, P. Epple,, W. F. Bibb,, R. M. McKinney,, R.W. Starnes,, J. M. Colville,, R. K. Selander,, P. A. Edelstein, and, C. W. Moss. 1988. Legionella pneumophila serogroup Lansing 3 isolated from a patient with fatal pneumonia, and descriptions of L. pneumophila subsp. pneumophila subsp. nov., L. pneumophila subsp. fraseri subsp. nov., and L. pneumophila subsp. pascullei subsp. nov. J. Clin. Microbiol. 26:16951703.
2. Fry, N. K.,, S. Alexiou-Daniel,, J. M. Bangsborg,, S. Bernander,, M. Castellani Pastoris,, J. Etienne,, M. Forsblom,, V. Gaia,, J. H. Helbig,, D. Lindsay,, 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. 5:462477.
3. Fry, N. K. and, T. G. Harrion. 1998. An evaluation of intergenic rRNA gene sequence length polymorphism analysis for the identification of Legionella species. J. Med. Microbiol. 47:667678.
4. 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. 43:20472052.
5. Helbig, J. H.,, S. Bernander,, M. Castellani Pastoris,, J. Etienne,, V. Gaia,, S. Lauwers,, D. Lindsay,, P. C. Lück,, T. Marques,, S. Mentula,, M. F. Peeters,, C. Pelaz,, M. Struelens,, S. A. Uldum,, G. Wewalka, and, T. G. Harrison (2002). Pan-European study on culture-proven Legionnaires’ disease: distribution of Legionella pneumophila serogroups and monoclonal subgroups. Eur. J. Clin. Microbiol Infect. Dis. 21:710716.
6. Helbig, J. H.,, J. B. Kurtz,, M. Castellani Pastoris,, C. Pelaz, and, P. C. Lück. 1997. The antigenic lipopolysaccharide components of Legionella pneumophila recognized by monoclonal antibodies: possibilities and limitations for division of the species into serogroups. J. Clin. Microbiol. 35:28412845.
7. Joly J. R.,, R. M. McKinney,, O. J. Tobin,, W. F. Bibb,, I. D. Watkins, and, D. Ramsay. 1986. Development of a standardized subgrouping scheme for Legionella pneumophila serogroup 1 using monoclonal antibodies. J. Clin. Microbiol. 23:768771.
8. Lück, P. C. and, J. H. Helbig. 2002. Typing of Legionella strains isolated from patients and environmental sources in Germany, 1990-2000, p. 267–270. In R. Marre et al. (ed.), Legionella. ASM Press, Washington, D.C.
9. Pringler, N.,, P. Brydov, and, S. A. Uldum. 2002. Occurrence of Legionella in Danish hot water systems, p.298301. In Legionella. R. Marre et al. (ed.), ASM Press, Washington, D.C.
10. Ratcliff, R. M.,, J. A. Lanser,, P. A. Manning, and, M. W. Heuzenroeder. 1998 Sequence-based classification scheme for the genus Legionella targeting the mip gene. J. Clin. Microbiol. 36:15601567.

Tables

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TABLE 1

Reaction pattern of reagents ( = 11) with respect to serogroup and monoclonal subgroup destination

Citation: Helbig J, Lück P. 2006. Serotyping of in Epidemiological Investigations: Limitations in the Era of Genotyping, p 68-72. In Cianciotto N, Kwaik Y, Edelstein P, Fields B, Geary D, Harrison T, Joseph C, Ratcliff R, Stout J, Swanson M (ed), . ASM Press, Washington, DC. doi: 10.1128/9781555815660.ch18

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