Epidemiology and Microbial Genomics, Laboratoire National de Santé, Dudelange, Luxembourg.
INRAE, Oniris, SECALIM, Nantes, France.
Front Cell Infect Microbiol. 2021 Jan 8;10:608020. doi: 10.3389/fcimb.2020.608020. eCollection 2020.
is the leading cause of bacterial gastroenteritis, which has motivated the monitoring of genetic profiles circulating in Luxembourg since 13 years. From our integrated surveillance using a genotyping strategy based on an extended MLST scheme including and markers, an unexpected endemic pattern was discovered in the temporal distribution of genotypes. We aimed to test the hypothesis of stable lineages occurrence by implementing whole genome sequencing (WGS) associated with comprehensive and internationally validated schemes. This pilot study assessed four WGS-based typing schemes to classify a panel of 108 strains previously identified as recurrent or sporadic profiles using this in-house typing system. The strain collection included four common lineages in human infection (N = 67) initially identified from recurrent combination of ST-- alleles also detected in non-human samples: veterinary (N = 19), food (N = 20), and environmental (N = 2) sources. An additional set of 19 strains belonging to sporadic profiles completed the tested panel. All the strains were processed by WGS by using Illumina technologies and by applying stringent criteria for filtering sequencing data; we ensure robustness in our genomic comparison. Four typing schemes were applied to classify the strains: (i) the cgMLST SeqSphere+ scheme of 637 loci, (ii) the cgMLST Oxford scheme of 1,343 loci, (iii) the cgMLST INNUENDO scheme of 678 loci, and (iv) the wgMLST INNUENDO scheme of 2,795 loci. A high concordance between the typing schemes was determined by comparing the calculated adjusted Wallace coefficients. After quality control and analyses with these four typing schemes, 60 strains were confirmed as members of the four recurrent lineages regardless of the method used (N = 32, 12, 7, and 9, respectively). Our results indicate that, regardless of the typing scheme used, epidemic or endemic signals were detected as reflected by lineage B (ST2254-9-1) in 2014 or lineage A (ST19-8-7), respectively. These findings support the clonal expansion of stable genomes in population exhibiting a multi-host profile and accounting for the majority of clinical strains isolated over a decade. Such recurring genotypes suggest persistence in reservoirs, sources or environment, emphasizing the need to investigate their survival strategy in greater depth.
是细菌性肠胃炎的主要病因,这促使卢森堡 13 年来一直在监测循环中的遗传特征。我们通过使用基于扩展 MLST 方案的基因分型策略进行综合监测,该方案包括 和 标记物,发现了基因型在时间分布上的意外地方性模式。我们旨在通过实施全基因组测序 (WGS) 并结合全面和国际上经过验证的方案来检验稳定谱系发生的假设。这项试点研究评估了四种基于 WGS 的分型方案,以对使用内部分型系统先前确定为反复或散发性图谱的 108 株菌株进行分类。该菌株集包括人类感染中的四个常见谱系(N = 67),最初是从非人类样本中检测到的反复出现的 ST--等位基因组合中识别出来的:兽医(N = 19)、食品(N = 20)和环境(N = 2)来源。另外一组 19 株属于散发性图谱的菌株完成了测试面板。所有菌株均通过 Illumina 技术进行 WGS 处理,并通过严格的过滤测序数据标准进行处理;我们确保基因组比较的稳健性。应用了四种分型方案对菌株进行分类:(i)637 个基因座的 cgMLST SeqSphere+方案,(ii)1343 个基因座的 cgMLST Oxford 方案,(iii)678 个基因座的 cgMLST INNUENDO 方案,以及(iv)2795 个基因座的 wgMLST INNUENDO 方案。通过比较计算出的调整 Wallace 系数,确定了分型方案之间的高度一致性。经过这四种分型方案的质量控制和分析,无论使用哪种方法,60 株均被确认为四个反复出现谱系的成员(分别为 N = 32、12、7 和 9)。我们的结果表明,无论使用哪种分型方案,都检测到了流行或地方性信号,这反映了 2014 年谱系 B(ST2254-9-1)或谱系 A(ST19-8-7)的情况。这些发现支持了在表现出多宿主特征并占过去十年分离的大多数临床菌株的 群体中稳定基因组的克隆扩张。这种反复出现的基因型表明在储层、来源或环境中存在持久性,强调需要更深入地研究其生存策略。