Phylogenetic analyses and detection of viridans streptococci based on sequences and denaturing gradient gel electrophoresis of the rod shape-determining protein gene.
Phylogenetic analyses and detection of viridans streptococci based on sequences and denaturing gradient gel electrophoresis of the rod shape-determining protein gene.
复制标题
DOI：
10.3402/jom.v1i0.2015
复制
发表时间：
2009-08-28
影响因子：
4.5
通讯作者：
Fujiwara, Taku
中科院分区：
文献类型：
--
作者：
Konishi, Ikuri;Hoshino, Tomonori;Fujiwara, Taku
背景：草绿色链球菌的群体分析非常重要，因为这些物种除了是人类口腔共生微生物群的重要成员之外，还与龋齿、菌血症和亚急性心内膜炎相关。设计：在这项研究中，我们对杆状决定蛋白基因（rodA）进行了系统发育分析，该基因与细胞形态、细胞分裂和抗生素敏感性相关，并证明 rodA 基因的多样性足以识别草绿色链球菌物种水平的链球菌。此外，我们还开发了一种基于rodA基因多样性的更方便的变性梯度凝胶电泳（DGGE）方法（rodA-DGGE），用于检测人类唾液中的9种优势链球菌，即血链球菌、口腔链球菌、轻链球菌、副血链球菌、戈登链球菌、前庭链球菌、唾液链球菌、变异链球菌和远亲链球菌。结果：事实证明，这种 rodA-DGGE 方法可用于检测草绿色链球菌，无需培养、分离和表型表征。结论：通过 rodA-DGGE 分析口腔微生物群比使用 16S rDNA 的传统 DGGE 分析具有更高的分辨率，并且可能成为口腔微生物诊断的替代方法。链球菌感染。
BACKGROUND: Population analysis of viridans streptococci is important because these species are associated with dental caries, bacteremia, and subacute endocarditis, in addition to being important members of the human oral commensal microbiota.DESIGN: In this study, we phylogenetically analyzed the rod shape-determining protein gene (rodA), which is associated with cellular morphology, cell division, and sensitivity for antibiotics, and demonstrated that the diversity of the rodA gene is sufficient to identify viridans streptococci at the species level. Moreover, we developed a more convenient denaturing gradient gel electrophoresis (DGGE) method based on the diversity of the rodA gene (rodA-DGGE) for detecting nine dominant streptococcal species in human saliva, namely, Streptococcus sanguinis, Streptococcus oralis, Streptococcus mitis, Streptococcus parasanguinis, Streptococcus gordonii, Streptococcus vestibularis, Streptococcus salivarius, Streptococcus mutans, and Streptococcus sobrinus.RESULTS: This rodA-DGGE method proved useful in detecting viridans streptococci without cultivation, isolation, and phenotypic characterization.CONCLUSION: Analysis of the oral microbiota by rodA-DGGE offers a higher resolution than the conventional DGGE using 16S rDNA and may be an alternative in the microbial diagnosis of streptococcal infection.