Houang E T, Chu Y, Ng T, Cheng A F
Department of Microbiology, Chinese University of Hong Kong, Prince of Wales Hospital, Hong Kong.
J Clin Microbiol. 1998 Sep;36(9):2404-7. doi: 10.1128/JCM.36.9.2404-2407.1998.
We used pulsed-field gel electrophoresis (PFGE) to study the genetic relatedness of 235 isolates of Shigella flexneri and Shigella sonnei collected in Hong Kong (97 isolates from 1986 and 1987 and 138 isolates from 1994 and 1995). Altogether, 13 gels were run with bacteriophage lambda ladder DNA (Pharmacia) as an external reference in every sixth lane, standardized reagents and methods, and isolates randomized for species and years. For quantitative illustration of the relationships within a large body of isolates, computer-generated dendrograms were used to determine the number of isolates in pulsotypes at Dice coefficients of similarity of 75% (PT75) and 50% (PT50). For S. flexneri, there was a significant difference in the distribution of isolates collected during the two periods in both PT75 and PT50, with 68% of isolates collected in 1994 and 1995 sharing a coefficient of similarity of >/=68%. For S. sonnei, a significant difference was observed in PT50 only. We also used Upholt's formula for an approximation of the fraction of nucleotide difference between isolates and Molecular Evolutionary Genetics Analysis to determine relative genetic distances. For both species, the relative genetic distances between isolates of the earlier collection period were significantly greater (P < 0.0001), i. e., they were further apart and therefore more diverse than those of the later period. We conclude that it is possible for a typical clinical laboratory to analyze a large amount of PFGE information on Shigella isolates obtained under controlled conditions. Such data analysis should enhance surveillance capabilities and give indications of further work to be done on various aspects of bacterial pathogenicity of the species.
我们采用脉冲场凝胶电泳(PFGE)技术,对在香港收集的235株福氏志贺菌和宋内志贺菌进行遗传相关性研究(其中97株分离自1986年和1987年,138株分离自1994年和1995年)。共进行了13次凝胶电泳,每隔六条泳道加入噬菌体λ阶梯DNA(Pharmacia公司产品)作为外部参照,使用标准化试剂和方法,并将分离株按菌种和年份随机排列。为了定量说明大量分离株之间的关系,利用计算机生成的树状图,以75%(PT75)和50%(PT50)的Dice相似系数来确定脉冲型中分离株的数量。对于福氏志贺菌,在PT75和PT50中,两个时期收集的分离株分布存在显著差异,1994年和1995年收集的分离株中有68%的相似系数≥68%。对于宋内志贺菌,仅在PT50中观察到显著差异。我们还使用Upholt公式估算分离株之间核苷酸差异的比例,并利用分子进化遗传学分析来确定相对遗传距离。对于这两个菌种,早期收集时期分离株之间的相对遗传距离显著更大(P<0.0001),即它们之间的距离更远,因此比后期收集的分离株更加多样。我们得出结论,典型的临床实验室有可能分析在受控条件下获得的大量志贺菌分离株PFGE信息。这样的数据分析应能增强监测能力,并为该菌种细菌致病性各个方面的进一步研究提供方向。