Zhang Jingyun, Diao Baowei, Zhang Na, Cui Zhigang, Zhang Lijuan, Xu Jianguo, Kan Biao
National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, P. O. Box 5, Changping, Beijing 102206, PR China.
J Microbiol Methods. 2007 Oct;71(1):15-22. doi: 10.1016/j.mimet.2007.07.002. Epub 2007 Jul 13.
Molecular subtyping is used to distinguish pathogenic bacterial strains during epidemiological surveys of infectious diseases and the discovery of novel pathogens. There is a need to standardize protocols involving Pulse-Field Gel Electrophoresis (PFGE), a crucial method in molecular subtyping, to make inter-laboratory results comparable and information exchange possible. The PFGE pattern varies with electrophoretic parameters (EPs), so it is important to select the parameter that can distinguish the patterns better. To optimize EPs of PFGE for Vibrio cholerae, we analyzed 24 isolates of V. cholerae O1 biotype El Tor and 26 isolates of O139 by PFGE with Not|. We used four different EPs and compared the similarity coefficients from the four groups by Friedman test. Based on the principle that the electrophoretic parameter producing smaller similarity in the PFGE patterns has higher level of discriminatory power, the one producing the smallest similarity coefficients is considered to be optimal. This method could be applied to determine the optimal molecular subtyping protocol for other bacteria.
在传染病流行病学调查和新型病原体发现过程中，分子分型用于区分致病细菌菌株。有必要对涉及脉冲场凝胶电泳（PFGE）的方案进行标准化，PFGE是分子分型中的关键方法，以便使实验室间的结果具有可比性并实现信息交流。PFGE图谱会因电泳参数（EPs）而有所不同，因此选择能更好区分图谱的参数很重要。为了优化霍乱弧菌PFGE的电泳参数，我们用NotI对24株霍乱弧菌O1生物型埃尔托生物型和26株O139菌株进行了PFGE分析。我们使用了四种不同的电泳参数，并通过弗里德曼检验比较了四组的相似系数。基于PFGE图谱中产生较小相似性的电泳参数具有更高鉴别力的原则，产生最小相似系数的参数被认为是最佳的。该方法可用于确定其他细菌的最佳分子分型方案。
