Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg, Denmark ; WHO Collaborating Centre for Antimicrobial Resistance in Food-borne Pathogens and EU Reference Laboratory for Antimicrobial Resistance, National Food Institute, Technical University of Denmark, Kgs. Lyngby, Denmark.
Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg, Denmark.
PLoS One. 2014 Jan 3;9(1):e84566. doi: 10.1371/journal.pone.0084566. eCollection 2014.
Multidrug-resistant Salmonella enterica serovar Typhimurium ST313 has emerged in sub-Saharan Africa causing severe infections in humans. Therefore, it has been speculated that this specific sequence type, ST313, carries factors associated with increased pathogenicity. We assessed the role in virulence of a gene with a yet unknown function, st313-td, detected in ST313 through comparative genomics. Additionally, the structure of the genomic island ST313-GI, harbouring the gene was determined. The gene st313-td was cloned into wild type S. Typhimurium 4/74 (4/74-C) as well as knocked out in S. Typhimurium ST313 02-03/002 (Δst313-td) followed by complementation (02-03/002-C). Δst313-td was less virulent in mice following i.p. challenge than the wild type and this phenotype could be partly complemented in trans, indicating that st313-td plays a role during systemic infection. The gene st313-td was shown not to affect invasion of cultured epithelial cells, while the absence of the gene significantly affects uptake and intracellular survival within macrophages. The gene st313-td was proven to be strongly associated to invasiveness, harboured by 92.5% of S. Typhimurium blood isolates (n = 82) and 100% of S. Dublin strains (n = 50) analysed. On the contrary, S. Typhimurium isolates of animal and food origin (n = 82) did not carry st313-td. Six human, non-blood isolates of S. Typhimurium from Belarus, China and Nepal harboured the gene and belonged to sequence types ST398 and ST19. Our data showed a global presence of the st313-td gene and in other sequence types than ST313. The gene st313-td was shown to be expressed during logarithmic phase of growth in 14 selected Salmonella strains carrying the gene. This study reveals that st313-td plays a role in S. Typhimurium ST313 pathogenesis and adds another chapter to understanding of the virulence of S. Typhimurium and in particular of the emerging sequence type ST313.
多重耐药沙门氏菌血清型肠炎亚种 ST313 已在撒哈拉以南非洲出现,导致人类严重感染。因此,人们推测这种特定的序列类型 ST313 携带与致病性增加相关的因素。我们通过比较基因组学评估了在 ST313 中检测到的一个具有未知功能的基因 st313-td 在毒力中的作用。此外,还确定了携带该基因的基因组岛 ST313-GI 的结构。将基因 st313-td 克隆到野生型鼠伤寒沙门氏菌 4/74(4/74-C)中,并在鼠伤寒沙门氏菌 ST313 02-03/002(Δst313-td)中敲除,然后进行互补(02-03/002-C)。与野生型相比,经腹腔感染后,Δst313-td 小鼠的毒力降低,该表型可部分在转染中互补,表明 st313-td 在全身感染过程中发挥作用。研究表明,该基因 st313-td 不影响培养的上皮细胞的侵袭,而该基因的缺失则显著影响巨噬细胞内的摄取和存活。研究证明,该基因 st313-td 与侵袭性密切相关,在所分析的 92.5%(n=82)的鼠伤寒沙门氏菌血分离株和 100%(n=50)的都柏林沙门氏菌株中均携带该基因。相反,动物和食品来源的鼠伤寒沙门氏菌分离株(n=82)不携带 st313-td。来自白俄罗斯、中国和尼泊尔的 6 株人非血源鼠伤寒沙门氏菌分离株携带该基因,属于序列类型 ST398 和 ST19。我们的数据显示,st313-td 基因在全球范围内存在,并且存在于除 ST313 以外的其他序列类型中。研究表明,在携带该基因的 14 株选定的沙门氏菌菌株中,该基因在对数生长期表达。这项研究表明,st313-td 基因在鼠伤寒沙门氏菌 ST313 发病机制中发挥作用,并为理解鼠伤寒沙门氏菌的毒力,特别是新兴的序列类型 ST313 的毒力增添了新的篇章。