African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12496

Full Length Research Paper

Multiplex polymerase chain reaction (PCR) assay for simultaneous detection of shiga-like toxin (stx1 and stx2), intimin (eae) and invasive plasmid antigen H (ipaH) genes in diarrheagenic Escherichia coli

Sharifi Yazdi MK1, Akbari A.2 and Soltan Dallal MM2,3*
  1Department of Medical Laboratory Sciences, Faculty of Paramedicene, Tehran University of Medical Sciences (TUMS), Tehran, Iran. 2Division of Microbiology, Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences (TUMS),  P.O. Box 14155-6446, Tehran, Iran. 3Research Microbial Resistance Center ,Tehran University of Medical Sciences.
Email: [email protected]

  •  Accepted: 07 December 2010
  •  Published: 28 February 2011

Abstract

 

Despite the fact that diarrheagenic Escherichia coli (DEC) has been identified as a major etiologic agent of diarrhea in children worldwide, few studies have beenperformed in Iran to evaluate the etiology of these organisms. To evaluate theetiology of shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC)and enteroinvasive E. coli (EIEC) in children with diarrhea in Iran a total 300 stool specimens from children with diarrhea were tested for the detection of E.coli. Out of 300 samples, 39 were identified as E. coli by biochemical tests and were subjected for serogrouping. The most prevalent serogroups among these isolates were serogroup IV, followed by III,I and II respectively. A single multiplex polymerase chain reaction (MPCR) was designed for the detection of target genesof stx1/ stx2, eae and ipaH in DEC. The dominating strain was EPEC (55.6%),followed by STEC (25%) and EIEC (19.4%).

 

Key words: Multiplex polymerase chain reaction (PCR), diarrheagenic Escherichia coli, shiga-like toxin.

Abbreviation

DEC, Diarrheagenic Escherichia coliEPEC, enteropathogenicEscherichia coliEHEC, enterohemorrhagic Escherichia coliEIEC,eenteroinvasive Escherichia coliETEC, enterotoxigenic Escherichia coliEAEC,enteroaggregative Escherichia coliDAEC, diffusely adherent Escherichia coli; LEE,  locus of enterocyte effacement; A/E, attaching and effacing; STEC, shiga toxin–producing Escherichia coliNCBI, National Center for Biotechnology Information; BLAST, Basic Local Alignment Search Tool; MPCR, multiplex polymerase chain reaction.