Nyholm Outi, Halkilahti Jani, Wiklund Gudrun, Okeke Uche, Paulin Lars, Auvinen Petri, Haukka Kaisa, Siitonen Anja
Bacterial Infections Unit, Department of Infectious Diseases, National Institute for Health and Welfare (THL), Helsinki, Finland.
Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.
PLoS One. 2015 Aug 27;10(8):e0135936. doi: 10.1371/journal.pone.0135936. eCollection 2015.
Shigatoxigenic Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC) cause serious foodborne infections in humans. These two pathogroups are defined based on the pathogroup-associated virulence genes: stx encoding Shiga toxin (Stx) for STEC and elt encoding heat-labile and/or est encoding heat-stable enterotoxin (ST) for ETEC. The study investigated the genomics of STEC/ETEC hybrid strains to determine their phylogenetic position among E. coli and to define the virulence genes they harbor.
The whole genomes of three STEC/ETEC strains possessing both stx and est genes were sequenced using PacBio RS sequencer. Two of the strains were isolated from the patients, one with hemolytic uremic syndrome, and one with diarrhea. The third strain was of bovine origin. Core genome analysis of the shared chromosomal genes and comparison with E. coli and Shigella spp. reference genomes was performed to determine the phylogenetic position of the STEC/ETEC strains. In addition, a set of virulence genes and ETEC colonization factors were extracted from the genomes. The production of Stx and ST were studied.
The human STEC/ETEC strains clustered with strains representing ETEC, STEC, enteroaggregative E. coli, and commensal and laboratory-adapted E. coli. However, the bovine STEC/ETEC strain formed a remote cluster with two STECs of bovine origin. All three STEC/ETEC strains harbored several other virulence genes, apart from stx and est, and lacked ETEC colonization factors. Two STEC/ETEC strains produced both toxins and one strain Stx only.
This study shows that pathogroup-associated virulence genes of different E. coli can co-exist in strains originating from different phylogenetic lineages. The possibility of virulence genes to be associated with several E. coli pathogroups should be taken into account in strain typing and in epidemiological surveillance. Development of novel hybrid E. coli strains may cause a new public health risk, which challenges the traditional diagnostics of E. coli infections.
产志贺毒素大肠杆菌(STEC)和产肠毒素大肠杆菌(ETEC)可导致人类严重的食源性感染。这两个致病菌群是根据与致病菌群相关的毒力基因来定义的:STEC的stx基因编码志贺毒素(Stx),ETEC的elt基因编码不耐热肠毒素和/或est基因编码耐热肠毒素(ST)。本研究调查了STEC/ETEC杂交菌株的基因组学,以确定它们在大肠杆菌中的系统发育位置,并确定它们所携带的毒力基因。
使用PacBio RS测序仪对三株同时携带stx和est基因的STEC/ETEC菌株的全基因组进行测序。其中两株菌株从患者中分离得到,一株来自溶血尿毒综合征患者,另一株来自腹泻患者。第三株菌株源自牛。对共享染色体基因进行核心基因组分析,并与大肠杆菌和志贺氏菌属参考基因组进行比较,以确定STEC/ETEC菌株的系统发育位置。此外,从基因组中提取了一组毒力基因和ETEC定植因子。研究了Stx和ST的产生情况。
人类STEC/ETEC菌株与代表ETEC、STEC、肠聚集性大肠杆菌以及共生和实验室适应性大肠杆菌的菌株聚类在一起。然而,牛源STEC/ETEC菌株与两株牛源STEC形成了一个较远的聚类。除stx和est外,所有三株STEC/ETEC菌株还携带其他几种毒力基因,并且缺乏ETEC定植因子。两株STEC/ETEC菌株都产生两种毒素,一株仅产生Stx。
本研究表明,不同大肠杆菌的致病菌群相关毒力基因可以共存于源自不同系统发育谱系的菌株中。在菌株分型和流行病学监测中应考虑毒力基因与多种大肠杆菌致病菌群相关的可能性。新型杂交大肠杆菌菌株的出现可能会带来新的公共卫生风险,这对传统的大肠杆菌感染诊断提出了挑战。