Optimization of Pulsed-Field Gel Electrophoresis for Legionella pneumophila Subtyping

Author:

Zhou Haijian1,Ren Hongyu1,Zhu Bingqing1,Kan Biao1,Xu Jianguo1,Shao Zhujun1

Affiliation:

1. National Institute for Communicable Disease Control and Prevention and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing, China

Abstract

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 .

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Cited by 14 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. Sequence-based typing of clinical and environmental Legionella pneumophila isolates in Shenyang, China;Enfermedades Infecciosas y Microbiología Clínica;2021-10

2. Sequence-based typing of clinical and environmental Legionella pneumophila isolates in Shenyang, China;Enfermedades infecciosas y microbiologia clinica (English ed.);2021-10

3. National Surveillance of Legionnaires’ Disease, China, 2014–2016;Emerging Infectious Diseases;2019-06

4. High Prevalence and Genetic Polymorphisms of Legionella in Natural and Man-Made Aquatic Environments in Wenzhou, China;International Journal of Environmental Research and Public Health;2017-02-24

5. Molecular epidemiology, phylogeny and evolution of Legionella;Infection, Genetics and Evolution;2016-09

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