Yamada-Noda Makiko, Ohkusu Kiyofumi, Hata Hiroyuki, Shah Mohammad Monir, Nhung Pham Hong, Sun Xiao Song, Hayashi Masahiro, Ezaki Takayuki
Department of Microbiology, Regeneration and Advanced Medical Science, Gifu University Graduate School of Medicine, Yanagido, Gifu, Japan.
Syst Appl Microbiol. 2007 Sep;30(6):453-62. doi: 10.1016/j.syapm.2007.06.003. Epub 2007 Jul 20.
The availability of the dnaJ1 gene for identifying Mycobacterium species was examined by analyzing the complete dnaJ1 sequences (approximately 1200 bp) of 56 species (54 of them were type strains) and comparing sequence homologies with those of the 16S rRNA gene and other housekeeping genes (rpoB, hsp65). Among the 56 Mycobacterium species, the mean sequence similarity of the dnaJ1 gene (80.4%) was significantly less than that of the 16S rRNA, rpoB and hsp65 genes (96.6%, 91.3% and 91.1%, respectively), indicating a high discriminatory power of the dnaJ1 gene. Seventy-one clinical isolates were correctly clustered to the corresponding type strains, showing isolates belonging to the same species. In order to propose a method for strain identification, we identified an area with a high degree of polymorphism, bordered by conserved sequences, that can be used as universal primers for PCR amplification and sequencing. The sequence of this fragment (approximately 350 bp) allows accurate species identification and may be used as a new tool for the identification of Mycobacterium species.
通过分析56个菌种（其中54个为模式菌株）的完整dnaJ1序列（约1200 bp），并将序列同源性与16S rRNA基因及其他管家基因（rpoB、hsp65）的序列同源性进行比较，研究了dnaJ1基因在分枝杆菌菌种鉴定中的可用性。在56个分枝杆菌菌种中，dnaJ1基因的平均序列相似性（80.4%）显著低于16S rRNA、rpoB和hsp65基因（分别为96.6%、91.3%和91.1%），表明dnaJ1基因具有较高的鉴别力。71株临床分离株被正确聚类到相应的模式菌株，表明属于同一菌种的分离株。为了提出一种菌株鉴定方法，我们鉴定了一个具有高度多态性的区域，其边界为保守序列，可作为PCR扩增和测序的通用引物。该片段（约350 bp）的序列能够准确鉴定菌种，可作为分枝杆菌菌种鉴定的新工具。
