Department of Microbiology & Immunology, Faculty of Tropical Medicine, Mahidol University, 420/6 Ratchawithi Road, Rajthevee District, Bangkok 10400, Thailand.
Infect Genet Evol. 2012 Oct;12(7):1368-76. doi: 10.1016/j.meegid.2012.03.025. Epub 2012 Apr 4.
Legionella-like organisms were isolated during 2003-2007 from various water resources by culturing on selective media of Wadowsky-Yee-Okuda agar. The 256 isolates were identified as belonging to the Legionella genus based on detection of 108 bp PCR product of the 5S rRNA gene, while the inclusion as Legionella pneumophila were confirmed by PCR detection of a specific mip gene region of 168 bp. The 50 isolates, identified as non-pneumophila, were then subjected to DNA tree analysis, based on mip gene of ~650 bp and rnpB genes product ranged from 304 to 354 bp. Phylogenetic tree was constructed to predict their species in relative to the available database. The isolates of which their speciation, based on those two genes were inconclusive, were then investigated for the almost full-length of 16S rRNA sequences. The isolates were assigned as 16 known Legionella species, and proposed seven novel species based on their unique 16S rRNA sequence.
2003-2007 年间，通过在 Wadowsky-Yee-Okuda 琼脂等选择性培养基上培养，从各种水源中分离出军团菌样生物体。根据 5S rRNA 基因的 108 bp PCR 产物检测，将 256 株分离物鉴定为军团菌属，而通过检测特定的 mip 基因区域 168 bp 的 PCR 检测来确认包含嗜肺军团菌。然后对 50 株鉴定为非嗜肺军团菌的分离物进行 DNA 树分析，基于~650 bp 的 mip 基因和 rnpB 基因产物范围为 304 至 354 bp。基于现有的数据库构建了系统发育树来预测它们的种属。基于这两个基因，对于那些种属分类不确定的分离物，进一步调查了其几乎全长的 16S rRNA 序列。将这些分离物分配到 16 种已知的军团菌种中，并根据其独特的 16S rRNA 序列提出了 7 种新的种。
