Chiou Chien-Shun, Watanabe Haruo, Wang You-Wun, Wang Wan-Ling, Terajima Jun, Thong Kwai-Lin, Phung Dac Cam, Tung Sheng Kai
Central Region Laboratory, Center for Research and Diagnostics, Centers for Disease Control, Taichung City, Taiwan.
J Clin Microbiol. 2009 Apr;47(4):1149-54. doi: 10.1128/JCM.01607-08. Epub 2009 Feb 18.
A panel of 916 isolates, including 703 closely related IST1 isolates, were characterized by inter-IS1 spacer typing (IST), pulsed-field gel electrophoresis (PFGE), and multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) to evaluate the utility of MLVA as a molecular tool for the phylogenetic analysis of Shigella sonnei. The global phylogenetic patterns determined by IST, PFGE, and MLVA were concordant. MLVA was carried out using 26 VNTR loci with a range of degrees of variability. MLVA data for the 703 IST1 isolates revealed that diversification among the closely related isolates was attributed mainly to four highly variable loci. The phylogenetic pattern for the closely related isolates determined using MLVA profiles of 8 highly variable loci was in agreement with that determined using the 26-locus profiles. A clustering analysis using the profiles of 18 loci with limited variability established clear phylogenetic relationships among IST clonal groups. Accordingly, MLVA is a useful tool for the phylogenetic analysis of S. sonnei. Combined VNTR loci with higher variability are useful markers for resolving closely related isolates, whereas combined loci with lower variability are suitable for establishing clear phylogenetic relationships between strains or clones that have evolved over a longer timescale.
一组916株分离株,包括703株密切相关的IST1分离株,通过IS1间隔区分型(IST)、脉冲场凝胶电泳(PFGE)和多位点可变数目串联重复序列(VNTR)分析(MLVA)进行特征分析,以评估MLVA作为宋内志贺氏菌系统发育分析分子工具的实用性。由IST、PFGE和MLVA确定的全球系统发育模式是一致的。MLVA使用26个具有不同变异程度的VNTR位点进行。703株IST1分离株的MLVA数据显示,密切相关分离株之间的多样性主要归因于四个高度可变的位点。使用8个高度可变位点的MLVA图谱确定的密切相关分离株的系统发育模式与使用26个位点图谱确定的模式一致。使用18个变异有限的位点的图谱进行聚类分析,在IST克隆组之间建立了明确的系统发育关系。因此,MLVA是宋内志贺氏菌系统发育分析的有用工具。具有较高变异性的组合VNTR位点是解析密切相关分离株的有用标记,而具有较低变异性的组合位点适合于在较长时间尺度上进化的菌株或克隆之间建立明确的系统发育关系。