Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Mar;76(5):1334-40.
doi: 10.1128/AEM.01455-09. Epub 2010 Jan 8.

Optimization of pulsed-field gel electrophoresis for Legionella pneumophila subtyping

Haijian Zhou  1 , Hongyu Ren, Bingqing Zhu, Biao Kan, Jianguo Xu, Zhujun Shao
Affiliations

Optimization of pulsed-field gel electrophoresis for Legionella pneumophila subtyping

Haijian Zhou et al. Appl Environ Microbiol. 2010 Mar.

Abstract

A total of 32 strains of Legionella pneumophila were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of L. pneumophila. Twenty-six isolates of L. pneumophila with various origins and 11 isolates from five different water systems were used as the panels. For optimization of electrophoretic parameters (EPs) of SfiI PFGE, 26 isolates were analyzed with SfiI digestion, using four EPs yielding the same D value. The EP of a switch time of 5 to 50 s for 21 h had the smallest similarity coefficients and was declared the optimal EP for SfiI PFGE of L. pneumophila. By software analysis and pilot study, AscI was chosen as another PFGE enzyme. AscI PFGE could cluster the isolates from each water system into the same or very similar patterns and had a high degree of typing concordance with other molecular methods. In evaluating the discriminatory power of AscI with the panel of 26 isolates, AscI PFGE gave one single pattern and a D value of 100%. AscI PFGE had a high discriminatory power and a high degree of consistency with epidemiological data and other molecular typing methods for L. pneumophila subtyping, and hence, AscI could be used as a restriction enzyme in PFGE subtyping of L. pneumophila.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Clustering results of patterns obtained using four EPs with SfiI digestion. Charts are shown for 26 L. pneumophila strains.
FIG. 2.
FIG. 2.
PFGE images of three L. pneumophila isolates restricted with AscI (lanes 2 to 4), RsrII (lanes 5 to 7), SgrAI (lanes 9 to 11), SbfI (lanes 13 to 15), SfiI (lanes 16 to 18), AsisI (lanes 20 to 22), and AvrII (lanes 24 to 26). The size standard (M) was loaded in lanes 1, 8, 12, 19, and 23.
FIG. 3.
FIG. 3.
Clustering results of patterns obtained with AscI digestion. Charts are shown for 26 L. pneumophila strains.
FIG. 4.
FIG. 4.
Clustering results of patterns obtained with SfiI digestion and AscI digestion of 11 L. pneumophila strains from five different water systems. The strains in each frame were from the same water system.
See this image and copyright information in PMC

References

    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. 49:1027-1033. - PubMed
    1. 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. - PMC - PubMed
    1. Blatt, S. P., M. D. Parkinson, E. Pace, P. Hoffman, D. Dolan, P. Lauderdale, R. A. Zajac, and G. P. Melcher. 1993. Nosocomial Legionnaires’ disease: aspiration as a primary mode of disease acquisition. Am. J. Med. 95:16-22. - PubMed
    1. Boccia, S., A. Stenico, R. Amore, L. Moroder, M. Orsini, V. Romano-Spica, and G. Ricciardi. 2005. Molecular epidemiology of Legionella pneumophila environmental isolates representing nine different serogroups determined by automated ribotyping and pulsed-field gel electrophoresis. Epidemiol. Infect. 133:1097-1105. - PMC - PubMed
    1. Breiman, R. F., W. Cozen, B. S. Fields, T. D. Mastro, S. J. Carr, J. S. Spika, and L. Mascola. 1990. Role of air sampling in investigation of an outbreak of Legionnaires’ disease associated with exposure to aerosols from an evaporative condenser. J. Infect. Dis. 161:1257-1261. - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources