J Bacteriol Virol.  2006 Sep;36(3):167-174. 10.4167/jbv.2006.36.3.167.

Genotypic and Phenotypic Characterization of Enteropathogenic Escherichia coli Isolated from Diarrheal Patients in Gwangju

• Kim SH
• Song HJ
• Chung JK
• Ha DR
• Ryu PY
• Lee JB

• KMID: 2055024
• DOI: http://doi.org/10.4167/jbv.2006.36.3.167

Abstract

Enteropathogenic Escherichia coli (EPEC) have been implicated in human diarrhea in several countries. Central to EPEC-mediated disease is its ability to cause intestinal lesions, known as attaching and effacing (A/E) lesion. We investigated 92 EPEC strains isolated from patients with diarrhea in Gwangju for their genotypic and phenotypic characteristics. Sixteen (17.4%) of all strains were found to be typical EPEC because they were bfpA gene positive by PCR. The most of typical EPEC isolates (87.5%) showed a localized adhesion (LA) pattern in Hep-2 cell adherence assay, whereas, only 11 atypical EPEC isolates (14.5%) were adhered to Hep-2 cells in a localized manner. Thirteen of the EPEC strains studied belonged to classical O-serogroups of EPEC and 7 isolates were classified as nonclassical EPEC serogroup and the other isolates could not be serotyped with our antisera. The subtypes of eae, tir, espA and espB genes which are major virulence genes concerned of A/E lesion on chromosome were analyzed by multiplex PCR for finding the original resource. The results showed that the composition of these genes subtypes was homogenous and heterogenous in 12 and 26 isolates, respectively. The others were non-determined type in terms of the gene subtype because of genetic diversity of intimin-coding eae genes. Our findings indicated that EPEC isolates from patients with diarrhea were diverse genetically and phenotypically, which require further study in regard to their virulence and epidemiological significance.

Keyword

EPEC; Attaching and effacing lesion; eae; bfpA; Localized adhesion (LA) pattern

MeSH Terms

Diarrhea
Enteropathogenic Escherichia coli*
Genetic Variation
Gwangju*
Humans
Immune Sera
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Virulence
Immune Sera

Figure

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  Figure 1. Adherence patterns in HEp-2 cells presented by EPEC isolates. A-1 and A-2, LA patterns (localized adherence). B-1 and B-2, AA patterns (aggregative adherence). C-1 and C-2, DA pattern (diffuse adherence). Original magnification, ×1000.

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  Figure 2. PCR result to detection of bfpA and EAF gene. A, lane 1: 100 bp ladder; lane 2: bfpA+ isolate; lane 3: bfpA– isolate. B, lane 1: 100 bp ladder; lane 2: EAF+ isolate; lane 3: EAF– isolate.

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  Figure 3. Result of multiplex PCR to analysis subtypes of eae, tir, espA and espB genes. A, lane 1: 100 bp ladder; lane 2: eaeα; lane 3: eaeβ; lane 4: eaeγ. B, lane 1: 100 bp ladder; lane 2: tirα: lane 3: tirβ; lane 4: tirγ; lane 5: negative control. C, lane 1: 100 bp ladder lane 2: espAα; lane 3: espAβ, lane 4: negative control. D, lane 1: 100 bp ladder; lane 2: espBα; lane 3: espBβ; lane 4: espBγ; lane 5: negative control.

Reference

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