College of Life Sciences, Qingdao University, Qingdao, P.R. China.
School of Medicine, Nankai University, Tianjin, P.R. China.
J Appl Microbiol. 2020 Sep;129(3):738-752. doi: 10.1111/jam.14637. Epub 2020 Mar 24.
To explore a prokaryotic species-specific DNA marker, 16S-23S rRNA gene internal transcribed spacer (ITS) sequence for identification and classification of Vibrio.
Five hundred and seventy four ITS sequences from 60 Vibrio strains were collected, then the primary and secondary structures of ITS sequence were analysed. The ITS was divided into several subunits, and the species-specificity of these subunits were evaluated by blast. The variable subunit of ITS showed high species-specificity. A protocol to identify a Vibrio species based on ITS analysis was developed and verified. Both the specificity and sensitivity were 100%. The phylogeny analysis of Vibrio based on ITS showed that ITS devised a better classification than 16S rDNA. Finally, an identification method of Vibrio based on ITS sequencing in food samples was developed and evaluated. The results of ITS sequencing were (100%) consistent with the results identified by ISO standard.
Vibrio could be accurately identified at the species level by using the ITS sequences.
The present study suggests that the ITS can be considered as a significant DNA marker for identification and classification of Vibrio species, and it posed a new path to screen the Vibrio in food sample.
探索一种原核生物种特异性 DNA 标记物，即 16S-23S rRNA 基因内转录间隔区（ITS）序列，用于鉴定和分类弧菌。
收集了 60 株弧菌的 574 个 ITS 序列，然后对 ITS 序列的一级和二级结构进行了分析。ITS 被分为几个亚单位，通过 blast 评估这些亚单位的种特异性。ITS 的可变亚单位表现出高度的种特异性。开发并验证了一种基于 ITS 分析鉴定弧菌种的方案，其特异性和灵敏度均为 100%。基于 ITS 的弧菌系统发育分析表明，ITS 比 16S rDNA 更能进行分类。最后，开发并评估了一种基于 ITS 测序鉴定食品样品中弧菌的方法。ITS 测序的结果与 ISO 标准鉴定的结果（100%）一致。
使用 ITS 序列可以准确鉴定弧菌的种属水平。
本研究表明，ITS 可作为鉴定和分类弧菌种的重要 DNA 标记物，为筛选食品样品中的弧菌提供了新途径。
