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Background A large outbreak of diarrhea and the hemolytic–uremic syndrome caused by an unusual serotype of Shiga-toxin–producing Escherichia coli (O104:H4) began in Germany in May 2011. As of July 22, a large number of cases of diarrhea caused by Shiga-toxin–producing E. Coli have been reported — 3167 without the hemolytic–uremic syndrome (16 deaths) and 908 with the hemolytic–uremic syndrome (34 deaths) — indicating that this strain is notably more virulent than most of the Shiga-toxin–producing E. Coli strains. Preliminary genetic characterization of the outbreak strain suggested that, unlike most of these strains, it should be classified within the enteroaggregative pathotype of E. Methods We used third-generation, single-molecule, real-time DNA sequencing to determine the complete genome sequence of the German outbreak strain, as well as the genome sequences of seven diarrhea-associated enteroaggregative E.
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DNA Preparation, Sequencing, and Comparative Analyses We purified and sequenced the genomic DNA from C227-11, from six African enteroaggregative E. Coli clinical isolates, and from five prototype enteroaggregative E. Coli isolates with the use of PacBio RS DNA sequencers (Sequence Read Archive accession number, SRA038239; GenBank accession number, AFST00000000). We also carried out a comparative analysis with the sequence of another outbreak isolate, TY2482. (See the article by Rohde et al. Elsewhere in this issue of the Journal for an analysis of sequence data obtained from TY2482. ) The provides additional details regarding DNA sequencing, assembly, resequencing analysis, and detection of DNA structural variations, as well as the construction of phylogenetic trees and the characterization of lambdalike phage elements, virulence factors, plasmids, and regions unique to the outbreak strain.

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In Panel A, the eight circular bands represent the tracks for the different enteroaggregative E. Coli (EAEC) O104:H4 isolates sequenced in our study (from inner to outer tracks: 55989, C227-11, C734-09, C35-10, C682-09, C760-09, C754-09, and C777-09). These different bands represent coverage plots mapped against the reference genome TY2482, with blue or green indicating a high degree of coverage, yellow moderate coverage, and orange or red little or no coverage. The C227-11 coverage plot indicates near-complete sequence identity with the TY2482 genome. The annotations on the outside of the chromosome plot highlight positions of known virulence-factor genes and genes previously characterized on the AA plasmid in the E. Coli O42 strain.

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Clin Infect Dis 20-1709|| • 10 Tompkins DS, Hudson MJ, Smith HR, et al. A study of infectious intestinal disease in England: microbiological findings in cases and controls. Commun Dis Public Health 1999;2:108-113[Erratum, Commun Dis Public Health 1999;2:222.] • 11 Okeke IN, Lamikanra A, Czeczulin J, Dubovsky F, Kaper JB, Nataro JP. Heterogeneous virulence of enteroaggregative Escherichia coli strains isolated from children in Southwest Nigeria.
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