Journal Information
Vol. 15. Issue 4.
Pages 365-369 (July - August 2011)
Share
Share
Download PDF
More article options
Vol. 15. Issue 4.
Pages 365-369 (July - August 2011)
Original article
Open Access
Comparison of multiplex PCR with serogrouping and PCR-RFLP of fliC gene for the detection of enteropathogenic Escherichia coli (EPEC)
Visits
3116
Saeid Bouzari1,
Corresponding author
saeidbouzari@yahoo.com

Correspondence to: Pasteur Institute of Iran, Molecular Biology Unit, Tehran, Iran 1316943551, Phone: +9821 66953311-20 (ext: 2223), Fax: +9821 66492619.
, Mohammad M. Aslani2, Mana Oloomi1, Anis Jafari1, Amir Dashti3
1 Researcher, Molecular Biology Unit, Pasteur Institute of Iran, Tehran, Iran
2 Researcher, Bacteriology Department, Pasteur Institute of Iran, Tehran, Iran
3 Researcher, Molecular Biology Unit, Pasteur Institute of Iran, Tehran, Iran
This item has received

Under a Creative Commons license
Article information
Abstract
Bibliography
Download PDF
Statistics
Abstract

Enteropathogenic Escherichia coli (EPEC) comprise one of the six categories of diarrhoeagenic E. coli (DEC). EPEC is subgrouped into typical (tEPEC) and atypical (aEPEC). The identification of DEC cannot be based only on cultural and biochemical criteria, since they are indistinguishable from the non-pathogenic E. coli commonly found in human feces. Several PCR methods, with both single and multiple target genes, have been reported for detecting the different DEC pathotypes. In the present study five hundred E. coli isolates from children with diarrhea were subjected into multiplex PCR. Furthermore the strains were typed serologically with O antisera and their fliC gene was characterized by PCR-RFLP. The results obtained revealed that overall 41 (8.2%) isolates could be detected as EPEC by this multiplex PCR assay. Of these isolates; 27 (66%) were typical (escv+, bfp+) and 14 (34%) atypical EPEC (escv+, bfp-). None of these 41 isolates contained the Stx1 and Stx2 genes. Among 37 (90%) typeable strains, nine different serogroups were present. The most common serogroups were O111, followed by O86, O55 and O119 and 10 different H types were found among these isolates. The multiplex PCR assay was found to be rapid and reliable in comparison to serological test; especially when screening the large number of isolates.

Keywords:
diarrhea
enteropathogenic Escherichia coli
polymerase chain reaction
Full text is only aviable in PDF
References
[1.]
R.M. Robins-Browne.
Traditional enteropathogenic Escherichia coli of infantile diarrhea.
Rev Infect Dis, 9 (1987), pp. 28-53
[2.]
L.R. Trabulsi, R. Keller, T.A. Tardelli Gomes.
Typical and atypical enteropathogenic Escherichia coli.
Emerg Infect Dis, 8 (2002), pp. 508-513
[3.]
H.W. Moon, S.C. Whipp, R.A. Argenzio, M.M. Levine, et al.
Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines.
Infect Immun, 41 (1983), pp. 1340-1351
[4.]
R.J. Rothbaum, J.C. Partin, K. Saalfield, et al.
An ultrastructural study of enteropathogenic Escherichia coli infection in human infants.
Ultrastruct Pathol, 4 (1983), pp. 291-304
[5.]
S. Tzipori, R.M. Robins-Browne, G. Gonis, et al.
Enteropathogenic Escherichia coli enteritis: evaluation of the gnotobiotic piglet as a model of human infection.
Gut, 26 (1985), pp. 570-578
[6.]
J. Celli, W. Deng, B.B. Finlay.
Enteropathogenic Escherichia coli (EPEC) attachment to epithelial cells: exploiting the host cell cytoskeleton from the outside.
Cell Microbiol, 2 (2000), pp. 1-9
[7.]
T. Tobe, T. Hayashi, C.G. Han, et al.
Complete DNA sequence and structural analysis of the enteropathogenic Escherichia coli adherence factor plasmid.
Infect Immun, 67 (1999), pp. 5455-5462
[8.]
J. Cleary, L.C. Lai, R.K. Shaw, et al.
Enteropathogenic Escherichia coli (EPEC) adhesion to intestinal epithelial cells: role of bundle-forming pili (BFP), EspA filaments and intimin.
Microbiology, 150 (2004), pp. 527-538
[9.]
T. Tobe, C. Sasakawa.
Role of bundle-forming pilus of enteropathogenic Escherichia coli in host cell adherence and in microcolony development.
Cell Microbiol, 3 (2001), pp. 579-585
[10.]
D. Bieber, S.W. Ramer, C.Y. Wu, et al.
Type IV pili, transient bacterial aggregates, and virulence of enteropathogenic Escherichia coli.
Science, 280 (1998), pp. 2114-2118
[11.]
M.S. Donnenberg, C.O. Tacket, S.P. James, et al.
Role of the eaeA gene in experimental enteropathogenic Escherichia coli infection.
J Clin Invest, 92 (1993), pp. 1412-1417
[12.]
Levine MM, Nataro JP, Karch H, et al. The diarrheal response of humans to some classic serotypes of enteropathogenic Escherichia coli is dependent on a plasmid encoding an enteroadhesiveness factor. J Infect Dis1985; 152(3):550-9.
[13.]
J. Antikainen, E. Tarkka, K. Haukka, et al.
New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples.
Eur J Clin Microbiol Infect Dis, 28 (2009), pp. 899-908
[14.]
K.R. Aranda, U. Fagundes-Neto, I.C. Scaletsky.
Evaluation of multiplex PCRs for diagnosis of infection with diarrheagenic Escherichia coli and Shigella spp.
J Clin Microbiol, 42 (2004), pp. 5849-5853
[15.]
L.T. Brandal, B.A. Lindstedt, L. Aas, et al.
Octaplex PCR and fluorescence-based capillary electrophoresis for identification of human diarrheagenic Escherichia coli and Shigella spp.
J Microbiol Methods, 68 (2007), pp. 331-341
[16.]
K. Kimata, T. Shima, M. Shimizu, et al.
Rapid categorization of pathogenic Escherichia coli by multiplex PCR.
Microbiol Immunol, 49 (2005), pp. 485-492
[17.]
M.A. Pass, R. Odedra, R.M. Batt.
Multiplex PCRs for identification of Escherichia coli virulence genes.
J Clin Microbiol, 38 (2000), pp. 2001-2004
[18.]
M. Vidal, E. Kruger, C. Durán, et al.
Single multiplex PCR assay to identify simultaneously the six categories of diarrheagenic Escherichia coli associated with enteric infections.
J Clin Microbiol, 43 (2005), pp. 5362-5365
[19.]
C. Hardegen, S. Messler, B. Henrich, et al.
A set of novel multiplex Taqman real-time PCRs for the detection of diarrhoeagenic Escherichia coli and its use in determining the prevalence of EPEC and EAEC in a university hospital.
Ann Clin Microbiol Antimicrob, 9 (2010), pp. 5-7
[20.]
D. Müller, P. Hagedorn, S. Brast, et al.
Rapid identification and differentiation of clinical isolates of enteropathogenic Escherichia coli (EPEC), atypical EPEC, and Shiga toxin-producing Escherichia coli by a one-step multiplex PCR method.
J Clin Microbiol, 44 (2006), pp. 2626-2629
[21.]
J. Sambrook, D.W. Russell.
Molecular cloning, Cold Spring Harbor Laboratory Press.
Cold Spring Harbor, (2001),
[22.]
R.J. Owen, P. Borman.
A rapid biochemical method for purifying high molecular weight bacterial chromosomal DNA for restriction enzyme analysis.
Nucleic Acids Res, 15 (1987), pp. 3631
[23.]
J. Machado, F. Grimont, P.A. Grimont.
Identification of Escherichia coli flagellar types by restriction of the amplified fliC gene.
Res Microbiol, 151 (2000), pp. 535-546
[24.]
World Health Organization Programme for Control of Diarrhoeal Diseases. Manual for Laboratory Investigations of Acute Enteric Infections. 1987; World Health Organization, Geneva.
[25.]
C.M. Abe, L.R. Trabulsi, J. Blanco, et al.
Virulence features of atypical enteropathogenic Escherichia coli identified by the eae(+) EAF-negative stx(-) genetic profile.
Diagn Microbiol Infect Dis, 64 (2009), pp. 357-365
[26.]
S.M. Tennant, M. Tauschek, K. Azzopardi, et al.
Characterization of atypical enteropathogenic E. coli strains of clinical origin.
BMC Microbiol, 9 (2009), pp. 117
[27.]
M.Y. Alikhani, A. Mirsalehian, M.M. Aslani.
Detection of typical and atypical enteropathogenic Escherichia coli (EPEC) in Iranian children with and without diarrhoea.
J Med Microbiol, 55 (2006), pp. 1159-1163
[28.]
S. Farooq, I. Hussain, M.A. Mir, et al.
Isolation of atypical enteropathogenic Escherichia coli and Shiga toxin 1 and 2f-producing Escherichia coli from avian species in India.
Lett Appl Microbiol, 48 (2009), pp. 692-697
Copyright © 2011. Elsevier Editora Ltda.. All rights reserved
Download PDF
The Brazilian Journal of Infectious Diseases
Article options
Tools