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Genetic profiling of enterohemorrhagic Escherichia coli strains in relation to clonality and clinical signs of infection.

ABSTRACT: Sixty-seven human strains of enterohemorrhagic Escherichia coli (EHEC) (from patients with more or less severe symptoms) were serogrouped and arranged according to pulsed-field gel electrophoresis (PFGE) patterns. We used PCR to investigate the strains according to known or putative virulence factors, and associations with disease were studied. All EHEC strains with the same PFGE pattern belonged to the same serogroup. On the contrary, two serogroups (O157 and O8) included strains with different PFGE patterns. We found several different combinations of chromosomal and plasmid-borne determinants, encoding the putative virulence factors, among the strains. As judged from clinical symptoms, there was no marked difference in pathogenicity among the strains and their combinations of virulence traits. All strains of O157 had the genes coding for verocytotoxin (VT) 2, intimin (eaeA), E. coli hemolysin (E-hly), and secreted serine protease (espP). Among EHEC non-O157 strains, the genes coding for VT1 and VT2 were equally dispersed. EaeA positivity was just as common among VT1- as VT2-positive strains. Among the plasmid-borne determinants, E-hly and espP were the most common and E-hly might be a pathogenicity marker among EHEC non-O157 strains. The conclusion is that PFGE is a very useful tool in epidemiological studies. The EHEC plasmids are heterogeneous in their gene composition, with the four plasmid-borne determinants found in many combinations. There was no reliable correlation between chromosomal and plasmid-borne virulence factors and human disease.

SUBMITTER: Welinder-Olsson C

PROVIDER: S-EPMC120270 | biostudies-literature | 2002 Mar

REPOSITORIES: biostudies-literature

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Genetic profiling of enterohemorrhagic Escherichia coli strains in relation to clonality and clinical signs of infection.

Welinder-Olsson Christina C Badenfors Maria M Cheasty Tom T Kjellin Eva E Kaijser Bertil B

Journal of clinical microbiology 20020301 3

Sixty-seven human strains of enterohemorrhagic Escherichia coli (EHEC) (from patients with more or less severe symptoms) were serogrouped and arranged according to pulsed-field gel electrophoresis (PFGE) patterns. We used PCR to investigate the strains according to known or putative virulence factors, and associations with disease were studied. All EHEC strains with the same PFGE pattern belonged to the same serogroup. On the contrary, two serogroups (O157 and O8) included strains with different ...[more]

PMID: 11880423

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Project description:Enterohemorrhagic E. coli (EHEC) is responsible for significant human illness, death, and economic loss. The main reservoir for EHEC is cattle, but plant-based foods are common vectors for human infection. Several outbreaks have been attributed to lettuce and leafy green vegetables grown in the Salinas and Santa Maria regions of California. Bacteria causing different outbreaks are mostly not close relatives, but one group of closely-related O157:H7 has caused several of them. This unusual pattern of recurrence may have some genetic basis. Here I use whole-genome sequences to reconstruct the genetic changes that occurred in the recent ancestry of this EHEC. In a short period of time corresponding to little genetic change, there were several changes to adhesion-related sequences, mainly adhesins. These changes may have greatly altered the adhesive properties of the bacteria. Possible consequences include increased persistence of cattle infections, more bacteria shed in cattle feces, and greater virulence in humans. Similar constellations of genetic change, which are detectable by current sequencing-based surveillance, may identify other bacteria that are particular threats to human health. In addition, the Santa Maria subclade carries a nonsense mutation affecting ArsR, a repressor of genes that confer resistance to arsenic and antimony. This suggests that the persistent source of Santa Maria contamination is located in an area with arsenic-contaminated groundwater, a problem in many parts of California. This inference may aid identification of the reservoir of EHEC, which would greatly aid mitigation efforts. IMPORTANCE Food-borne bacterial infections cause substantial illness and death. Understanding how bacteria contaminate food and cause disease is important for combating the problem. Closely-related E. coli, likely originating in cattle, have repeatedly caused outbreaks spread by vegetables grown in California. Such recurrence is atypical, and might have a genetic basis. The genetic changes that occurred in the recent ancestry of these E. coli can be reconstructed from their DNA sequences. Several mutations affect genes involved in bacterial adhesion. These might affect persistence of infection in cattle, quantity of bacteria in their feces, and human disease. They also suggest a way of detecting dangerous bacteria from their genome sequences. Furthermore, a subgroup carries a mutation affecting the regulation of genes conferring arsenic resistance. This suggests that the reservoir for contamination utilizes groundwater contaminated with arsenic, a problem in parts of California. This observation may be an aid to locating the persistent reservoir of contamination.

Project description:BackgroundEnterohaemorrhagic E. coli (EHEC) can cause severe disease such as bloody diarrhoea and haemolytic uraemic syndrome in humans. Besides production of Shiga toxins, the presence of LEE (eae-gene) and non-LEE (nle) encoded effector genes harboured on O-islands OI-122, OI-71 and OI-57 is associated with EHEC virulence and their frequency in outbreaks. Genes encoded by the EHEC-plasmid are putative virulence markers of EHEC. EHEC-plasmids, LEE and non-LEE effector genes have also been detected in some strains of enteropathogenic E. coli (EPEC). The objective of this study was to analyze the relationship between EHEC and EPEC for virulence genes encoded by genomic O-islands and by the EHEC-plasmids.ResultsNle genes ent/espL2, nleB and nleE (OI-122), nleA, nleF and nleH1-2 (OI-71), nleG5-2 and nleG6-2 (OI-57), espK (CP-933N) and the EHEC-plasmid encoded genes ehxA, espP, etpD and katP were searched in 73 typical and in 235 atypical enteropathogenic E. coli (EPEC) strains. Typical and atypical EPEC each fall into two clusters. Cluster 1 typical (n = 46) and atypical (n = 129) EPEC strains were characterized by the presence of OI-122 encoded genes and grouped together with 64 investigated EHEC strains. Cluster 2 typical (n = 27) and atypical (n = 106) strains grouped together with 52 LEE-negative, Shiga toxin-producing E. coli (STEC) and with 21 apathogenic E. coli strains. Typical EPEC Cluster 1 strains belonged to serotypes frequently involved in severe illness and outbreaks in children (O111:H2, O114:H2, O55:H6, O127:H6 and O142:H6). Atypical EPEC Cluster 1 strains were characterized by serotypes related to EHEC (O26:H11, O55:H7, O145:H28, O103:H2 and O103:H25).ConclusionThe OI-122 encoded nleB gene was found to be most closely associated with Cluster 1 strains and may serve as a diagnostic tool for the identification of virulent EHEC and EPEC seropathotypes. OI-71 encoded genes nleA, nleF and nleH1-2 are less associated with Cluster 1 strains. EHEC-plasmid, OI-57 and CP-933 associated genes showed only weak similarities with virulent Cluster 1 EHEC and EPEC strains.

Dataset Information

Genetic profiling of enterohemorrhagic Escherichia coli strains in relation to clonality and clinical signs of infection.

Publications

Genetic profiling of enterohemorrhagic Escherichia coli strains in relation to clonality and clinical signs of infection.

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