Chattaway Marie A, Schaefer Ulf, Tewolde Rediat, Dallman Timothy J, Jenkins Claire
Gastrointestinal Bacteria Reference Unit and Bioinformatics Unit, National Infection Service, Public Health England, London, United Kingdom.
Gastrointestinal Bacteria Reference Unit and Bioinformatics Unit, National Infection Service, Public Health England, London, United Kingdom
J Clin Microbiol. 2017 Feb;55(2):616-623. doi: 10.1128/JCM.01790-16. Epub 2016 Dec 14.
Escherichia coli and Shigella species are closely related and genetically constitute the same species. Differentiating between these two pathogens and accurately identifying the four species of Shigella are therefore challenging. The organism-specific bioinformatics whole-genome sequencing (WGS) typing pipelines at Public Health England are dependent on the initial identification of the bacterial species by use of a kmer-based approach. Of the 1,982 Escherichia coli and Shigella sp. isolates analyzed in this study, 1,957 (98.4%) had concordant results by both traditional biochemistry and serology (TB&S) and the kmer identification (ID) derived from the WGS data. Of the 25 mismatches identified, 10 were enteroinvasive E. coli isolates that were misidentified as Shigella flexneri or S. boydii by the kmer ID, and 8 were S. flexneri isolates misidentified by TB&S as S. boydii due to nonfunctional S. flexneri O antigen biosynthesis genes. Analysis of the population structure based on multilocus sequence typing (MLST) data derived from the WGS data showed that the remaining discrepant results belonged to clonal complex 288 (CC288), comprising both S. boydii and S. dysenteriae strains. Mismatches between the TB&S and kmer ID results were explained by the close phylogenetic relationship between the two species and were resolved with reference to the MLST data. Shigella can be differentiated from E. coli and accurately identified to the species level by use of kmer comparisons and MLST. Analysis of the WGS data provided explanations for the discordant results between TB&S and WGS data, revealed the true phylogenetic relationships between different species of Shigella, and identified emerging pathoadapted lineages.
大肠杆菌和志贺氏菌属密切相关,在遗传学上属于同一物种。因此,区分这两种病原体并准确鉴定志贺氏菌的四个物种具有挑战性。英国公共卫生部门针对特定生物体的生物信息学全基因组测序(WGS)分型流程依赖于使用基于kmer的方法对细菌物种进行初步鉴定。在本研究分析的1982株大肠杆菌和志贺氏菌属分离株中,1957株(98.4%)通过传统生物化学和血清学方法(TB&S)以及从WGS数据得出的kmer鉴定(ID)获得了一致的结果。在鉴定出的25个不匹配结果中,有10个是肠侵袭性大肠杆菌分离株,通过kmer ID被误鉴定为福氏志贺菌或鲍氏志贺菌,还有8个福氏志贺菌分离株由于福氏志贺菌O抗原生物合成基因无功能,被TB&S误鉴定为鲍氏志贺菌。基于从WGS数据得出的多位点序列分型(MLST)数据对种群结构进行分析表明,其余不一致的结果属于克隆复合体288(CC288),包括鲍氏志贺菌和痢疾志贺菌菌株。TB&S和kmer ID结果之间的不匹配是由于这两个物种之间密切的系统发育关系所致,并通过参考MLST数据得以解决。通过使用kmer比较和MLST,可以将志贺氏菌与大肠杆菌区分开来,并准确鉴定到物种水平。对WGS数据的分析为TB&S和WGS数据之间的不一致结果提供了解释,揭示了不同志贺氏菌物种之间真正的系统发育关系,并鉴定出新兴的致病适应谱系。