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16S rDNA分析与rep-PCR基因组指纹技术在假结核耶尔森菌分子鉴定中的比较

Comparison of 16S rDNA analysis and rep-PCR genomic fingerprinting for molecular identification of Yersinia pseudotuberculosis.

作者信息

Kim Wonyong, Song Mi-Ok, Song Wonkeun, Kim Ki-Jung, Chung Sang-In, Choi Chul-Soon, Park Yong-Ha

机构信息

Department of Microbiology, Chung-Ang University College of Medicine, Seoul 156-756, Korea.

出版信息

Antonie Van Leeuwenhoek. 2003;83(2):125-33. doi: 10.1023/a:1023301924932.

Abstract

16S rDNA sequence analysis and repetitive element sequence-based PCR (rep-PCR) genomic fingerprinting were evaluated on 11 type strains of the genus Yersinia and 17 recognized serotype strains of Y. pseudotuberculosis to investigate their genetic relatedness and to establish the value of techniques for the identification of Y. pseudotuberculosis. A phylogenetic tree constructed from 16S rDNA sequences showed that the type strains of Yersinia species formed distinct clusters with the exception of Y. pestis and Y. pseudotuberculosis. Moreover, Y. pestis NCTC 5923T was found to be closely related to Y. pseudotuberculosis serotypes 1b, 3, and 7. Dendrograms generated from REP-PCR, and ERIC-PCR data revealed that members of the genus Yersinia differed from each other with the degree of similarity 62% and 58%, respectively. However, the BOX-PCR results showed that Y. pestis 5923T clustered with the Y. pseudotuberculosis group with a degree of similarity 74%. According to these findings, 16S rDNA sequence analysis was unable to reliably discriminate Y. pseudotuberculosis from Y. pestis. However, REP-PCR and especially ERIC-PCR provided an effective means of differentiating between members of the taxa.

摘要

对11株耶尔森氏菌属模式菌株和17株已确认的假结核耶尔森氏菌血清型菌株进行了16S rDNA序列分析和基于重复元件序列的PCR(rep-PCR)基因组指纹分析,以研究它们的遗传相关性,并确定这些技术在鉴定假结核耶尔森氏菌方面的价值。由16S rDNA序列构建的系统发育树表明,除了鼠疫耶尔森氏菌和假结核耶尔森氏菌外,耶尔森氏菌属的模式菌株形成了不同的簇。此外,发现鼠疫耶尔森氏菌NCTC 5923T与假结核耶尔森氏菌血清型1b、3和7密切相关。从REP-PCR和ERIC-PCR数据生成的树状图显示,耶尔森氏菌属成员彼此不同,相似程度分别为62%和58%。然而,BOX-PCR结果表明,鼠疫耶尔森氏菌5923T与假结核耶尔森氏菌群聚集在一起,相似程度为74%。根据这些发现,16S rDNA序列分析无法可靠地区分假结核耶尔森氏菌和鼠疫耶尔森氏菌。然而,REP-PCR尤其是ERIC-PCR提供了一种区分分类单元成员的有效方法。

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