J Bacteriol Virol.  2007 Jun;37(2):69-78. 10.4167/jbv.2007.37.2.69.

Pathotypic Characterization of Enterocyte Effacement-related LEE Genes in EHEC and EPEC Isolated from Diarrheal Patients

Affiliations
  • 1Department of Microbiology and Immunology, College of Medicine, Pusan National University, Pusan 602-739, Korea. ybkim@pusan.ac.kr

Abstract

Attaching and effacing Escherichia coli (AEEC) cause enteric infections in humans and animals. Attaching indicates the intimate attachment of bacteria to the enterocyte, and effacing relates to the localized effacement of brush border microvilli. Enteropathogenic (EPEC) and enterohemorrhagic Escherichia coli (EHEC) infections are characterized by the formation of attaching and effacing (AE) lesion on the intestinal epithelial cells. Therefore, they are often grouped together as AEEC. Development of multiplex PCR allowed us to type five of the most important genes implicated in the formation of the AE lesion. A total of 60 AEEC strains isolated from diarrheal patients were investigated by multiplex PCR for the presence of the insertion site of locus of enterocyte effacement (LEE) and LEE-related (eae, tir, espA, espB, and espD) genes. Associating the results of LEE genes typing in the AEEC strains, three different pathotypes are determined: eae(gamma)-tir(gamma)-espA(gamma)-espB(gamma)-espD(gamma) (O157:H7), eae(beta)-tir(beta)-espA(beta)-espB(beta)-espD(beta) (O26:H11), and eae(alpha)-tir(alpha)-espA(alpha)-espB(alpha)-espD(alpha) (O55:H6). These results indicate that AEEC are a heterogenous groups of organisms.

Keyword

AEEC; LEE; Attaching and effacing; Multiplex PCR

MeSH Terms

Animals
Bacteria
Enterocytes*
Enterohemorrhagic Escherichia coli*
Enteropathogenic Escherichia coli*
Epithelial Cells
Escherichia coli
Humans
Microvilli
Multiplex Polymerase Chain Reaction

Figure

  • Figure 1. stx genes obtained from AEEC strains by multiplex PCR A∼C. A: lane M, 100 bp plus ladder (molecular weight markers); lane 1, control strains EDL933 (stx1+, stx2+); lane 2, EHEC O1157:H– strain (stx1–, stx2+); lane 3, EHEC O111:H– strain (stx1+, stx2–); lane 4, EPEC O55:H– strain (stx1–, stx2–). B: lane N, 100 bp ladder (molecular weight markers); lane 1, E. coli EDL933 (stx2e–, stx2f +); lane 2, EHEC O1157:H–strain (stx2e–, stx2f+).

  • Figure 2. Multiplex PCR to distinguish an intact selC locus from one disrupted by LEE. Primers K255 and K260 are predicted to produce a 418 bp amplicon in strains containing a LEE-disrupted locus. Primers K261 and K260 produced the 527 bp amplicon in strains indicating an intact selC locus. Lane M, 100 bp plus ladder (molecular weight markers); lane 1, E. coli EDL933 (stx1+, stx2+, control strains); lane 2, EHEC strain (O111:H–).


Reference

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