Emond-Rheault Jean-Guillaume, Hamel Jérémie, Jeukens Julie, Freschi Luca, Kukavica-Ibrulj Irena, Boyle Brian, Tamber Sandeep, Malo Danielle, Franz Eelco, Burnett Elton, Daigle France, Arya Gitanjali, Sanderson Kenneth, Wiedmann Martin, Slawson Robin M, Weadge Joel T, Stephan Roger, Bekal Sadjia, Gruenheid Samantha, Goodridge Lawrence D, Levesque Roger C
Institut de Biologie Intégrative et des Systèmes (IBIS), Université Laval, Québec, QC G1V 0A6, Canada.
Health Canada, Bureau of Microbial Hazards, Ottawa, ON K1A 0K9, Canada.
Microorganisms. 2020 Jul 8;8(7):1016. doi: 10.3390/microorganisms8071016.
The emergence of multidrug-resistant bacterial strains worldwide has become a serious problem for public health over recent decades. The increase in antimicrobial resistance has been expanding via plasmids as mobile genetic elements encoding antimicrobial resistance (AMR) genes that are transferred vertically and horizontally. This study focuses on , one of the leading foodborne pathogens in industrialized countries. S. enterica is known to carry several plasmids involved not only in virulence but also in AMR. In the current paper, we present an integrated strategy to detect plasmid scaffolds in whole genome sequencing (WGS) assemblies. We developed a two-step procedure to predict plasmids based on i) the presence of essential elements for plasmid replication and mobility, as well as ii) sequence similarity to a reference plasmid. Next, to confirm the accuracy of the prediction in 1750 short-read sequencing data, we combined Oxford Nanopore MinION long-read sequencing with Illumina MiSeq short-read sequencing in hybrid assemblies for 84 isolates to evaluate the proportion of plasmid that has been detected. At least one scaffold with an origin of replication (ORI) was predicted in 61.3% of the isolates tested. The results indicated that IncFII and IncI1 ORIs were distributed in many serotypes and were the most prevalent AMR genes carrier, whereas IncHI2A/IncHI2 and IncA/C2 were more serotype restricted but bore several AMR genes. Comparison between hybrid and short-read assemblies revealed that 81.1% of plasmids were found in the short-read sequencing using our pipeline. Through this process, we established that plasmids are prevalent in and we also substantially expand the AMR genes in the resistome of this species.
近几十年来,全球范围内多重耐药细菌菌株的出现已成为公共卫生领域的一个严重问题。抗菌药物耐药性的增加通过质粒作为编码抗菌药物耐药性(AMR)基因的移动遗传元件在垂直和水平方向上传播。本研究聚焦于工业化国家主要的食源性病原体之一。肠炎沙门氏菌已知携带多种质粒,这些质粒不仅与毒力有关,还与AMR有关。在本文中,我们提出了一种在全基因组测序(WGS)组装中检测质粒支架的综合策略。我们开发了一个两步程序来预测质粒,该程序基于:i)质粒复制和移动所必需元件的存在,以及ii)与参考质粒的序列相似性。接下来,为了在1750个短读长测序数据中确认预测的准确性,我们将牛津纳米孔MinION长读长测序与Illumina MiSeq短读长测序相结合,对84个分离株进行混合组装,以评估已检测到的质粒比例。在61.3%的测试分离株中预测到至少一个带有复制起点(ORI)的支架。结果表明,IncFII和IncI1 ORI分布在许多血清型中,是最普遍的AMR基因载体,而IncHI2A/IncHI2和IncA/C2的血清型限制更大,但携带多个AMR基因。混合组装和短读长组装之间的比较表明,使用我们的流程在短读长测序中发现了81.1%的质粒。通过这个过程,我们确定质粒在肠炎沙门氏菌中普遍存在,并且我们还大幅扩展了该物种耐药基因组中的AMR基因。