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Chapter 43 : Pulsed-Field Gel Electrophoresis Analysis and Sequence-Based Typing of Serogroup 1 Isolates from Japan

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Pulsed-Field Gel Electrophoresis Analysis and Sequence-Based Typing of Serogroup 1 Isolates from Japan, Page 1 of 2

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Abstract:

Pulsed-field gel electrophoresis (PFGE) is considered to be one of the most discriminative epidemiological methods for subtyping strains and for elucidating the sources of infection. On the other hand, sequence-based typing (SBT), which was recently developed, is also a powerful epidemiological method. All serogroup 1 isolates used in a study were independently obtained from a wide variety of Japanese locations. The primers used for SBT and the reaction mixture and conditions were the same as those used by Gaia et al. Putative novel variants found in this study were submitted to the curators of the European Working Group for Legionella Infections SBT database for verification and assignment of new allelic numbers according to the curators’ instructions. Both PFGE and SBT indicated that the spa-bath isolates from Japan were highly diverse. The water used in public spa baths in Japanese resorts is mostly obtained from hot springs. The first clinical isolate in Japan was not assigned to the cooling tower SB-type. Only four SB types were common between Europe and Japan. Although PFGE is the most widely used technique and is generally accepted to be highly effective in discriminating genomic differences, it may have certain drawbacks with regard to interlaboratory reproducibility. SBT appears to be less effective at discriminating between strains than PFGE. The authors have to consider the advantages and limitations of both methods and apply the most suitable method according to the requirements.

Citation: Amemura-Maekawa J, Kura F, Chang B, Watanabe H. 2006. Pulsed-Field Gel Electrophoresis Analysis and Sequence-Based Typing of Serogroup 1 Isolates from Japan, p 159-162. 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.ch43

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Pulsed-Field Gel Electrophoresis
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Legionella pneumophila
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Figures

Image of FIGURE 1
FIGURE 1

Isolates of serogroup 1 in Japan were used in this study. All isolates are epidemiologically unrelated to each other. The names of these isolates are indicated on the map along with the names of the prefectures where they were isolated. C, isolates from cooling tower water; B, isolates from spa and/or hot spring bath water; P, isolates from patients assumed to be infected at spas and/or hot spring baths; P*, isolates from patients infected at unknown sources; BP, representative isolates obtained from both patients and bath water in an outbreak. The P isolates are indicated on the prefectures where the patients visited, and P* isolates are indicated on the prefectures where the hospitals of patients’ admission were located.

Citation: Amemura-Maekawa J, Kura F, Chang B, Watanabe H. 2006. Pulsed-Field Gel Electrophoresis Analysis and Sequence-Based Typing of Serogroup 1 Isolates from Japan, p 159-162. 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.ch43
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Image of FIGURE 2
FIGURE 2

Cluster dendrogram by PFGE and sequence-based types of serogroup 1 isolates in Japan. Allele numbers are according to the European Working Group for Infections SBT database for (http://www.ewgli.org.), and new alleles that were determined in this research are underlined. Characteristics of hot spring water were quoted from the reference of the hot spring.

Citation: Amemura-Maekawa J, Kura F, Chang B, Watanabe H. 2006. Pulsed-Field Gel Electrophoresis Analysis and Sequence-Based Typing of Serogroup 1 Isolates from Japan, p 159-162. 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.ch43
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References

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1. Amemura-Maekawa, J.,, F. Kura,, B. Chang, and, H. Watanabe. 2005. Legionella pneumophila serogroup 1 isolates from cooling towers in Japan form a distinct genetic cluster. Microbiol. Immunol. 49:10271033.
2. Amemura-Maekawa, J.,, F. Kura,, H. Watanabe,, F. Gondaira, and, J. Sugiyama. 2002. Analysis of Legionella pneumophila serogroup 1 isolates in Japan by using pulsed-field gel elec-trophoresis and monoclonal antibodies, p. 302304. In R. Marre,,Y.Abu Kwaik, C. Bartlett,, N. P. Cianciotto,, B. S. Fields,, M. Frosch,, J. Hacker, and P. C. Lück (ed.). Legionella, ASM Press, Washington, D.C.
3. De Zoysa, A. S., and, T. G. Harrison. 1999. Molecular typing of Legionella pneumophila serogroup 1 by pulsed-field gel electrophoresis with SfiI and comparison of this method with restriction fragment-length polymorphism analysis. J. Med. Microbiol. 48:269278.
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. Gaia, V.,, N. K. Fry,, T. G. Harrison, and, R. Gaia, V.,, N. K. Peduzzi. 2003. Sequence-based typing of Legionella pneumophila serogroup 1 offers the potential for true portability in legionellosis outbreak investigation. J. Clin. Microbiol. 41:29322939.
6. Saito, A.,, T. Shimoda,, M. Nagasawa,, H. Tanaka,, N. Ito,, Y. Shigeno,, K. Yamaguchi,, M. Hirota,, M. Nakatomi, and, K. Hara. 1981.The first case of Legionnaires’ disease in Japan. Kansensyogaku Zasshi 55:124128.

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