Higgins Paul G, Prior Karola, Harmsen Dag, Seifert Harald
Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany.
German Centre for Infection Research (DZIF), partner site Bonn-Cologne, Cologne, Germany.
PLoS One. 2017 Jun 8;12(6):e0179228. doi: 10.1371/journal.pone.0179228. eCollection 2017.
We have employed whole genome sequencing to define and evaluate a core genome multilocus sequence typing (cgMLST) scheme for Acinetobacter baumannii. To define a core genome we downloaded a total of 1,573 putative A. baumannii genomes from NCBI as well as representative isolates belonging to the eight previously described international A. baumannii clonal lineages. The core genome was then employed against a total of fifty-three carbapenem-resistant A. baumannii isolates that were previously typed by PFGE and linked to hospital outbreaks in eight German cities. We defined a core genome of 2,390 genes of which an average 98.4% were called successfully from 1,339 A. baumannii genomes, while Acinetobacter nosocomialis, Acinetobacter pittii, and Acinetobacter calcoaceticus resulted in 71.2%, 33.3%, and 23.2% good targets, respectively. When tested against the previously identified outbreak strains, we found good correlation between PFGE and cgMLST clustering, with 0-8 allelic differences within a pulsotype, and 40-2,166 differences between pulsotypes. The highest number of allelic differences was between the isolates representing the international clones. This typing scheme was highly discriminatory and identified separate A. baumannii outbreaks. Moreover, because a standardised cgMLST nomenclature is used, the system will allow inter-laboratory exchange of data.
我们运用全基因组测序来定义和评估鲍曼不动杆菌的核心基因组多位点序列分型(cgMLST)方案。为定义核心基因组,我们从NCBI下载了总共1573个推测的鲍曼不动杆菌基因组以及属于先前描述的8个国际鲍曼不动杆菌克隆谱系的代表性分离株。然后将核心基因组应用于总共53株耐碳青霉烯鲍曼不动杆菌分离株,这些分离株先前通过脉冲场凝胶电泳(PFGE)分型,并与德国8个城市的医院暴发相关联。我们定义了一个由2390个基因组成的核心基因组,其中平均98.4%的基因能从1339个鲍曼不动杆菌基因组中成功检出,而医院不动杆菌、皮氏不动杆菌和醋酸钙不动杆菌的良好靶标率分别为71.2%、33.3%和23.2%。在对先前鉴定的暴发菌株进行测试时,我们发现PFGE和cgMLST聚类之间具有良好的相关性,在一个脉冲型内等位基因差异为0 - 8个,在不同脉冲型之间差异为40 - 2166个。等位基因差异数量最多的是代表国际克隆的分离株之间。这种分型方案具有高度鉴别力,能识别出不同的鲍曼不动杆菌暴发。此外,由于使用了标准化的cgMLST命名法,该系统将允许实验室间的数据交换。