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基于 16S-23S rDNA 内转录间隔区的 DNA 标记对链球菌进行种特异性鉴定。

Species-Specific Identification of Streptococcus based on DNA Marker in 16S-23S rDNA Internal Transcribed Spacer.

机构信息

College of Life Sciences, Qingdao University, Qingdao, People's Republic of China.

School of Medicine, Nankai University, Tianjin, People's Republic of China.

出版信息

Curr Microbiol. 2020 Aug;77(8):1569-1579. doi: 10.1007/s00284-020-01975-8. Epub 2020 Apr 6.

Abstract

Streptococcus is closely correspondent to human. The accurate species-specific identification method of Streptococcus is important for the bacteria clinical diagnosis, molecular epidemiological analysis, and microecological study. In the last decades, DNA markers are widely utilized for identification of prokaryotic species. However, 16S rDNA, the most popular bacterial DNA marker, cannot properly distinguish closely related Streptococcus species. In present study, we employed 16S-23S rRNA gene internal transcribed spacer (ITS) sequence to explore the species-specific DNA marker. We predicted the secondary structure of Streptococcus ITS sequence transcribed products. Then we identified that the specific and consensus sequences in the primary structure can be found occupying an individual subunit in the secondary structure, which explained the foundation of the mosaic-like structure of ITS. We evaluated the specificity of ITS in Streptococcus, and found that the specificity can be detected by a further analysis of a BLAST result. Then, we developed an identification procedure based on the ITS sequence. We verified the procedure by 500 ITS sequence. The accuracy rate of this procedure was 100% for Streptococcus at genus level, and 99.3% at species level. It suggested that ITS can be utilized to accurately identify Streptococcus at the species level. This work suggests that further exploration of ITS could be applied in other bacterial genera for identification and classification, which may be a useful topic for future microbiology studies.

摘要

链球菌与人密切相关。链球菌的准确种特异性鉴定方法对于细菌临床诊断、分子流行病学分析和微生态学研究非常重要。在过去的几十年中,DNA 标记物广泛用于鉴定原核生物物种。然而,16S rDNA,最流行的细菌 DNA 标记物,不能正确地区分密切相关的链球菌种。在本研究中,我们采用 16S-23S rRNA 基因内转录间隔区(ITS)序列来探索种特异性 DNA 标记物。我们预测了链球菌 ITS 序列转录产物的二级结构。然后,我们发现,一级结构中的特异性和保守序列可以在二级结构中占据单个亚基,这解释了 ITS 镶嵌结构的基础。我们评估了 ITS 在链球菌中的特异性,并发现通过进一步分析 BLAST 结果可以检测到特异性。然后,我们基于 ITS 序列开发了一种鉴定程序。我们通过 500 个 ITS 序列验证了该程序。该程序在属水平上对链球菌的准确率为 100%,在种水平上为 99.3%。这表明 ITS 可用于准确鉴定链球菌种。这项工作表明,进一步探索 ITS 可应用于其他细菌属的鉴定和分类,这可能是未来微生物学研究的一个有用课题。

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