2Centre for Systems Genomics, University of Melbourne, Parkville, Victoria 3010, Australia.
1Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia.
Microb Genom. 2016 Jul 11;2(7):e000064. doi: 10.1099/mgen.0.000064. eCollection 2016 Jul.
The lipopolysaccharide (O) and flagellar (H) surface antigens of are targets for serotyping that have traditionally been used to identify pathogenic lineages. These surface antigens are important for the survival of within mammalian hosts. However, traditional serotyping has several limitations, and public health reference laboratories are increasingly moving towards whole genome sequencing (WGS) to characterize bacterial isolates. Here we present a method to rapidly and accurately serotype isolates from raw, short read WGS data. Our approach bypasses the need for genome assembly by directly screening WGS reads against a curated database of alleles linked to known and novel O-groups and H-types (the EcOH database) using the software package srst2. We validated the approach by comparing results for 197 enteropathogenic isolates with those obtained by serological phenotyping in an independent laboratory. We then demonstrated the utility of our method to characterize isolates in public health and clinical settings, and to explore the genetic diversity of >1500 genomes from multiple sources. Importantly, we showed that transfer of O- and H-antigen loci between chromosomal backbones is common, with little evidence of constraints by host or pathotype, suggesting that strain space' may be virtually unlimited, even within specific pathotypes. Our findings show that serotyping is most useful when used in combination with strain genotyping to characterize microevolution events within an inferred population structure.
脂多糖 (O) 和鞭毛 (H) 表面抗原是血清分型的目标,传统上用于识别病原谱系。这些表面抗原对于 在哺乳动物宿主中的生存至关重要。然而,传统的血清分型有几个局限性,公共卫生参考实验室越来越倾向于使用全基因组测序 (WGS) 来描述细菌分离株。在这里,我们提出了一种从原始短读 WGS 数据中快速准确地对 分离株进行血清分型的方法。我们的方法通过使用软件包 srst2 直接将 WGS 读数与已知和新型 O 群和 H 型相关联的等位基因的精心整理的数据库(EcOH 数据库)进行筛选,绕过了 基因组组装的需要。我们通过将 197 个肠致病性 分离株的 结果与独立实验室进行的血清表型学比较来验证该方法。然后,我们展示了我们的方法在公共卫生和临床环境中对分离株进行特征描述的实用性,并探索了来自多个来源的 >1500 个基因组的遗传多样性。重要的是,我们表明 O-和 H-抗原基因座在 染色体骨架之间的转移很常见,宿主或病原体类型的限制很少,这表明“菌株空间”可能几乎是无限的,即使在特定病原体类型中也是如此。我们的研究结果表明,血清分型最有用的是与菌株基因分型结合使用,以描述推断的种群结构内的微观进化事件。