Application of rpoB and Zinc Protease Gene for Use in Molecular Discrimination of Fusobacterium nucleatum Subspecies
Application of rpoB and Zinc Protease Gene for Use in Molecular Discrimination of Fusobacterium nucleatum Subspecies
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DOI：
10.1128/jcm.01631-09
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发表时间：
2010-02-01
影响因子：
9.4
通讯作者：
Kook, Joong-Ki
中科院分区：
文献类型：
--
作者：
Kim, Hwa-Sook;Lee, Dae-Sil;Kook, Joong-Ki
有核梭杆菌分为5个亚种，栖息在人类口腔（具核梭杆菌亚种）。具核体；多形菌；梭形虫，具核孢子虫亚种。和具核孢子虫亚种。基于几种表型特征和DNA-DNA杂交模式。然而，在亚种水平上检测或鉴别临床分离的具核梭菌的方法是费力的，昂贵的，耗时的。因此，本研究通过分析RNA聚合酶β亚基基因（rpoB）和锌蛋白酶基因的核苷酸序列来区分核仁赤豆的亚种。测定并分析了5个亚种、28株临床分离的具核f菌和10株作为对照组的牙周f菌的rpoB基因（约2419 bp）、锌蛋白酶基因（878 bp）和16S rRNA基因（约1500 bp）的部分序列。系统发育数据表明，rpoB和锌蛋白酶基因序列清晰地描述了核仁赤豆的亚种，比16S rRNA基因序列在这方面提供了更高的分辨率。根据rpoB和锌蛋白酶基因的系统发育分析，核仁F. nucleatum亚种。青霉属和具核菌属亚种。梭形虫可归为单一亚种。5个临床分离株可作为具核梭菌的新亚种。结果表明，rpoB和锌蛋白酶基因是核仁梭菌亚种鉴别和分类分析的有效靶点。
Fusobacterium nucleatum is classified into five subspecies that inhabit the human oral cavity (F. nucleatum subsp. nucleatum, F. nucleatum subsp. polymorphum, F. nucleatum subsp. fusiforme, F. nucleatum subsp. vincentii, and F. nucleatum subsp. animalis) based on several phenotypic characteristics and DNA-DNA hybridization patterns. However, the methods for detecting or discriminating the clinical isolates of F. nucleatum at the subspecies levels are laborious, expensive, and time-consuming. Therefore, in this study, the nucleotide sequences of the RNA polymerase beta-subunit gene (rpoB) and zinc protease gene were analyzed to discriminate the subspecies of F. nucleatum. The partial sequences of rpoB (approximately 2,419 bp), the zinc protease gene (878 bp), and 16S rRNA genes (approximately 1,500 bp) of the type strains of five subspecies, 28 clinical isolates of F. nucleatum, and 10 strains of F. periodonticum (as a control group) were determined and analyzed. The phylogenetic data showed that the rpoB and zinc protease gene sequences clearly delineated the subspecies of F. nucleatum and provided higher resolution than the 16S rRNA gene sequences in this respect. According to the phylogenetic analysis of rpoB and the zinc protease gene, F. nucleatum subsp. vincentii and F. nucleatum subsp. fusiforme might be classified into a single subspecies. Five clinical isolates could be delineated as a new subspecies of F. nucleatum. The results suggest that rpoB and the zinc protease gene are efficient targets for the discrimination and taxonomic analysis of the subspecies of F. nucleatum.