Wang Min, Cao Boyang, Yu Qunfang, Liu Lei, Gao Qili, Wang Lei, Feng Lu
TEDA School of Biological Sciences and Biotechnology, Nankai University, 23 HongDa Street, TEDA, Tianjin 300457, China.
J Clin Microbiol. 2008 Nov;46(11):3555-63. doi: 10.1128/JCM.00927-08. Epub 2008 Aug 27.
The 16S-23S rRNA gene internal transcribed spacer (ITS) regions of Klebsiella spp., including Klebsiella pneumoniae subsp. pneumoniae, Klebsiella pneumoniae subsp. ozaenae, Klebsiella pneumoniae subsp. rhinoscleromatis, Klebsiella oxytoca, Klebsiella planticola, Klebsiella terrigena, and Klebsiella ornithinolytica, were characterized, and the feasibility of using ITS sequences to discriminate Klebsiella species and subspecies was explored. A total of 336 ITS sequences from 21 representative strains and 11 clinical isolates of Klebsiella were sequenced and analyzed. Three distinct ITS types-ITS(none) (without tRNA genes), ITS(glu) [with a tRNA(Glu (UUC)) gene], and ITS(ile+ala) [with tRNA(Ile (GAU)) and tRNA(Ala (UGC)) genes]-were detected in all species except for K. pneumoniae subsp. rhinoscleromatis, which has only ITS(glu) and ITS(ile+ala). The presence of ITS(none) in Enterobacteriaceae had never been reported before. Both the length and the sequence of each ITS type are highly conserved within the species, with identity levels from 0.961 to 1.000 for ITS(none), from 0.967 to 1.000 for ITS(glu), and from 0.968 to 1.000 for ITS(ile+ala). Interspecies sequence identities range from 0.775 to 0.989 for ITS(none), from 0.798 to 0.997 for ITS(glu), and from 0.712 to 0.985 for ITS(ile+ala). Regions with significant interspecies variations but low intraspecies polymorphisms were identified; these may be targeted in the design of probes for the identification of Klebsiella to the species level. Phylogenetic analysis based on ITS regions reveals the relationships among Klebsiella species similarly to that based on 16S rRNA genes.
对肺炎克雷伯菌亚种肺炎克雷伯菌、肺炎克雷伯菌亚种臭鼻克雷伯菌、肺炎克雷伯菌亚种鼻硬结克雷伯菌、产酸克雷伯菌、植生克雷伯菌、土生克雷伯菌和解鸟氨酸克雷伯菌等克雷伯菌属细菌的16S - 23S rRNA基因内部转录间隔区（ITS）进行了特征分析，并探讨了利用ITS序列区分克雷伯菌属种和亚种的可行性。对来自21株代表性菌株和11株克雷伯菌临床分离株的336条ITS序列进行了测序和分析。除鼻硬结克雷伯菌亚种仅具有ITS（glu）和ITS（ile + ala）外，在所有种中均检测到三种不同的ITS类型——ITS（无）（无tRNA基因）、ITS（glu）[带有tRNA（Glu（UUC））基因]和ITS（ile + ala）[带有tRNA（Ile（GAU））和tRNA（Ala（UGC））基因]。肠杆菌科中ITS（无）的存在此前从未有过报道。每种ITS类型的长度和序列在种内高度保守，ITS（无）的同一性水平为0.961至1.000，ITS（glu）为0.967至1.000，ITS（ile + ala）为0.968至1.000。种间序列同一性ITS（无）为0.775至0.989，ITS（glu）为0.798至0.997，ITS（ile + ala）为0.712至0.985。确定了种间差异显著但种内多态性低的区域；这些区域可作为设计用于克雷伯菌属种水平鉴定探针的靶点。基于ITS区域的系统发育分析揭示了克雷伯菌属种之间的关系，与基于16S rRNA基因的分析相似。
