Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 52: 192-201, 2006. First published December 29, 2005; 10.1373/clinchem.2005.059766
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Data Supplements
Right arrow All Versions of this Article:
clinchem.2005.059766v1
52/2/192    most recent
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (17)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Garrido, P.
Right arrow Articles by Parro, V.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Garrido, P.
Right arrow Articles by Parro, V.
Related Collections
Right arrow Molecular Diagnostics and Genetics
(Clinical Chemistry. 2006;52:192-201.)
© 2006 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

STEC-EPEC Oligonucleotide Microarray: A New Tool for Typing Genetic Variants of the LEE Pathogenicity Island of Human and Animal Shiga Toxin–Producing Escherichia coli (STEC) and Enteropathogenic E. coli (EPEC) Strains

Patricia Garrido1, Miguel Blanco2, Mercedes Moreno-Paz1, Carlos Briones1, Ghizlane Dahbi2, Jesús Blanco2, Jorge Blanco2 and Víctor Parro1,a

1 Laboratorio de Ecología Molecular, Centro de Astrobiología (INTA-CSIC), Madrid, Spain.
2 Laboratorio de Referencia de E. coli (LREC), Departamento de Microbiología y Parasitología, Facultad de Veterinaria, Universidad de Santiago de Compostela, Lugo, Spain.

aAddress correspondence to this author at: Laboratorio de Ecología Molecular, Centro de Astrobiología (INTA-CSIC), carretera de Ajalvir km 4, 28850, Torrejón de Ardoz, Madrid, Spain. Fax 34-915201074; e-mail parrogv{at}inta.es.

Background: Shiga toxin–producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) are important emerging pathogens that can cause a severe and sometimes fatal illness. Differentiation of eae, tir, espA, espD, and espB gene variants of the locus of enterocyte effacement (LEE) pathogenicity island represents an important tool for typing in routine diagnostics as well as in pathogenesis, epidemiologic, clonal, and immunologic studies.

Methods: Type-specific oligonucleotide microarrays and a PCR scheme were designed and constructed for the detection and typing of genetic variants of the LEE genes. Oligonucleotide probes were tested for their specificity against the corresponding type strain by microarray hybridization using fluorescent DNA, either PCR-amplified (single, multiplex, long-range), chromosomal, or amplified chromosomal DNA.

Results: The PCR scheme and the oligonucleotide microarray allowed us to distinguish 16 variants ({alpha}1, {alpha}2, ß1, ß2, {gamma}1, {gamma}2/{theta}, {delta}/{kappa}, {epsilon}, {zeta}, {eta}, {iota}, {lambda}, µ, {nu}, {xi}, o) of the eae gene, 4 variants ({alpha}1, ß1, {gamma}1, {gamma}2/{theta}) of the tir gene, 4 variants ({alpha}1, ß1, ß2, {gamma}1) of the espA gene, 3 variants ({alpha}1, ß1, {gamma}1) of the espB gene, and 3 variants ({alpha}1, ß1, {gamma}1) of the espD gene. We found a total of 12 different combinations of tir, espA, espB, and espD genes among the 25 typed strains.

Conclusions: The PCR scheme and the oligonucleotide microarray described are effective tools to rapidly screen multiple virulence genes and their variants in E. coli strains isolated from human and animal infections. The results demonstrate the great genetic diversity among LEE genes of human and animal STEC and EPEC strains.




The following articles in journals at HighWire Press have cited this article:


Home page
 Vet Rec.Home page
Y. Ha, S. K. Ha, Y. H. Lee, D. Kim, S. Chae, C. H. Kim, K. K. Ahn, O. Kim, and C. Chae
Prevalence of tir gene subtypes in Escherichia coli isolates from pigs
Vet Rec., October 4, 2008; 163(14): 424 - 425.
[Full Text] [PDF]

Home page
 J. Clin. Microbiol.Home page
J. E. Afset, E. Anderssen, G. Bruant, J. Harel, L. Wieler, and K. Bergh
Phylogenetic Backgrounds and Virulence Profiles of Atypical Enteropathogenic Escherichia coli Strains from a Case-Control Study Using Multilocus Sequence Typing and DNA Microarray Analysis
J. Clin. Microbiol., July 1, 2008; 46(7): 2280 - 2290.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Microbiol.Home page
K. Ito, M. Iida, M. Yamazaki, K. Moriya, S. Moroishi, J. Yatsuyanagi, T. Kurazono, N. Hiruta, and O.-A. Ratchtrachenchai
Intimin Types Determined by Heteroduplex Mobility Assay of Intimin Gene (eae)-Positive Escherichia coli Strains
J. Clin. Microbiol., March 1, 2007; 45(3): 1038 - 1041.
[Abstract] [Full Text] [PDF]

Home page
 J Med MicrobiolHome page
M. Blanco, J. E. Blanco, G. Dahbi, A. Mora, M. P. Alonso, G. Varela, M. P. Gadea, F. Schelotto, E. A. Gonzalez, and J. Blanco
Typing of intimin (eae) genes from enteropathogenic Escherichia coli (EPEC) isolated from children with diarrhoea in Montevideo, Uruguay: identification of two novel intimin variants ({micro}B and {xi}R/{beta}2B).
J. Med. Microbiol., September 1, 2006; 55(Pt 9): 1165 - 1174.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2006 by the American Association for Clinical Chemistry.