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采用脉冲场凝胶电泳对变形链球菌分离株进行DNA指纹分析。

DNA fingerprinting of isolates of Streptococcus mutans by pulsed-field gel electrophoresis.

作者信息

Mineyama R, Yoshino S, Maeda N

机构信息

Department of Oral Microbiology, School of Dentistry at Niigata, Nippon Dental University, 1-8 Hamauracho, 951-8580 Niigata, Japan.

出版信息

Microbiol Res. 2007;162(3):244-9. doi: 10.1016/j.micres.2006.06.014. Epub 2006 Jul 25.

Abstract

Forty isolates and five standard laboratory strains, representing serotypes c, e and f of Streptococcus mutans were analyzed by pulsed-field gel electrophoresis (PFGE) after digestion of the genomic DNA with BssH II. The digestion patterns of standard laboratory strains were characteristic of serotypes c, e and f. Serotypes c and f generated diagnostic DNA fragments of approximately 145 kbp and of approximately 130-175 kbp in length, respectively. Serotype e generated a ladder of at least 14 fragments of 15-155 kbp in length. The digestion patterns of isolates were essentially similar to those of the standard laboratory strains. The patterns of almost all isolates obtained from a single individual were identical, but patterns of a few different types were also observed among isolates obtained from two individuals. Digestion with BssH II revealed differences among isolates obtained from different individuals. We used differences in banding patterns among isolates to construct a dendrogram. The dendrogram included two major clusters, one that consisted of isolates of serotypes c and f, and an other that consisted of isolates of serotype e. Our results indicate that BssH II is a useful enzyme for distinguishing among isolates of S. mutans and that digestion patterns obtained by PFGE can be used for chromosomal DNA fingerprinting.

摘要

用BssH II消化基因组DNA后,通过脉冲场凝胶电泳(PFGE)分析了代表变形链球菌血清型c、e和f的40株分离株和5株标准实验室菌株。标准实验室菌株的消化模式具有血清型c、e和f的特征。血清型c和f分别产生长度约为145 kbp和约130 - 175 kbp的诊断性DNA片段。血清型e产生了至少14个长度为15 - 155 kbp片段的梯状条带。分离株的消化模式与标准实验室菌株基本相似。从单个个体获得的几乎所有分离株的模式都是相同的,但在从两个个体获得的分离株中也观察到了一些不同类型的模式。用BssH II消化揭示了从不同个体获得的分离株之间的差异。我们利用分离株之间条带模式的差异构建了一个树状图。该树状图包括两个主要聚类,一个由血清型c和f的分离株组成,另一个由血清型e的分离株组成。我们的结果表明,BssH II是一种用于区分变形链球菌分离株的有用酶,并且通过PFGE获得的消化模式可用于染色体DNA指纹分析。

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