Department of Respiratory Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, P.O. Box 5, Changping, Beijing 102206, People's Republic of China.
Appl Environ Microbiol. 2010 Mar;76(5):1334-40. doi: 10.1128/AEM.01455-09. Epub 2010 Jan 8.
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.
总共使用了 32 株嗜肺军团菌菌株来优化脉冲场凝胶电泳（PFGE），以对嗜肺军团菌进行分型。26 株来自不同来源的嗜肺军团菌分离株和来自五个不同水系统的 11 株分离株被用作面板。为了优化 SfiI PFGE 的电泳参数（EP），用 SfiI 消化分析了 26 株分离株，使用了产生相同 D 值的四个 EP。切换时间为 5 到 50 s 的 EP 具有最小的相似系数，并被宣布为嗜肺军团菌 SfiI PFGE 的最佳 EP。通过软件分析和初步研究，选择了 AscI 作为另一种 PFGE 酶。AscI PFGE 可以将来自每个水系统的分离株聚类到相同或非常相似的模式中，并且与其他分子方法具有高度的分型一致性。在评估 26 株分离株的 AscI 区分能力时，AscI PFGE 给出了一个单一的模式和 100%的 D 值。AscI PFGE 具有高的区分能力和与流行病学数据和其他分子分型方法的高度一致性，因此，AscI 可用于嗜肺军团菌的 PFGE 分型。
