Article Figures & Data
Figures
- Fig. 1.
Multiplex PCR analysis of fecal cultures from HUS patients and STEC isolates. Crude DNA extracts of broth cultures from the indicated samples or strains were analyzed by multiplex PCR assay 1, as previously described (19). PCR products were electrophoresed on 2% agarose gels and stained with ethidium bromide. Lanes: M, DNA size markers (pUC19 DNA digested with HpaII; fragment sizes visible are 501/489, 404, 331, 242, 190, and 147 bp); 1, negative control; 2, positive control (O111:H− STEC strain 95NR1, which is positive for stx1,stx2, eae, and EHEC hlyA); 3, fecal culture from HUS patient 1; 4, fecal culture from HUS patient 2; 5, fecal culture from HUS patient 3; 6, STEC isolate 98NK2; 7, STEC isolate 98BN1. The expected mobilities for the various specific PCR products are also indicated.
- Fig. 2.
Western immunoblot detection of anti-O113 LPS. Aliquots of LPS purified from E. coli serogroups O113 (lanes 1), O111 (lanes 2), and O157 (lanes 3) were separated by SDS-PAGE, transferred to nitrocellulose filters, and reacted with convalescent-phase sera from the three HUS patients or serum from a healthy control.
- Fig. 3.
RFLP analysis of STEC O113 isolates. Genomic DNA purified from the indicated STEC strains was digested withEcoRI (A) or SphI (B), electrophoresed, and subjected to Southern hybridization analysis with anstx2-specific DNA probe, as described previously (24). Lanes: 1, 98NK2; 2, 98BN1; 3, 97MW1; 4, MW10; 5, 1183; 6, 3848. Lane M contains DNA size markers of 23.1, 9.4, 6.6, 4.4, 2.3, and 2.0 kb.
- Fig. 4.
Pulsed-field gel electrophoresis ofXbaI-digested genomic DNA of STEC O113. Lanes 1 to 6 are labelled as for Fig. 3. The lanes marked M contain marker DNA (SmaI-digested genomic DNA of Staphylococcus aureus NCTC8325).
- Fig. 5.
Deduced amino acid sequences of the A and B subunits of Stx2O113 aligned with published sequences for Stx2 (10), Stx2c (27), and Stx2d (9). Dots denote residues identical to Stx2O113; the dash denotes an absent residue. Arrows indicate the first residue of the mature polypeptide for each subunit.
