Submitted: 27 Nov 2013
Revised: 15 Dec 2013
Accepted: 24 Dec 2013
First published online: 05 Oct 2016
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Int J Enteric Pathog. 2014;2(1): e16431.
doi: 10.17795/ijep16431
  Abstract View: 2778
  PDF Download: 1883

Research Article

Multiplex Real-Time PCR Assay for the Detection of LT, STIa and STIb Genes in Enterotoxigenic Escherichia coli

Pejman Abbasi 1, Mohammad Kargar 2 * , Abbas Doosti 3, Jalal Mardaneh 1, Mohammad Ali Dehyadegari 1, Sadegh Ghorbani-Dalini 4

1 Alborzi Clinical Microbiology Research Center, Namazee Hospital, Shiraz University of Medical Sciences, Shiraz, I.R. Iran
2 Department of Microbiology, Islamic Azad University, Jahrom, I.R. Iran
3 Biotechnology Research Center, Islamic Azad University, Shahrekord Branch, Shahrekord, I.R. Iran
4 Department of Biology, University of Sistan and Baluchistan, Zahedan, I.R. Iran
*Corresponding author: Mohammad Kargar, Department of Microbiology, Islamic Azad University, Jahrom, I.R. Iran. Tel: +98-9177164249, Email:


Background: Diarrhea is an important cause of illness and death among all age groups on a global scale. Enterotoxigenic Escherichia coli (ETEC) protozoa were established as a causative agent of diarrhea in developing and developed countries. The identification of diarrheagenic E. coli (DEC) strains needs to detect the factors that determine the virulence of these organisms.

Objectives: In this study, we aimed to use the multiplex real-time (MRT) and multiplex PCR assays for identification of ETEC in patients with diarrhea in Shiraz, Iran.

Patients and Methods: A total of 430 stool samples were collected from patients with diarrhea in Shiraz, in 2012. Diarrheagenic E. coli (DEC) strains were isolated by standard biochemical analysis. We used MRT-PCR and multiplex PCR (MPCR) assays to detect the presence of LT, STΙa and STΙb genes in ETEC.

Results: In this study, 430 stool samples were tested and 52 (12.1%) were identified as contaminated with E. coli using standard biochemical tests. The ETEC were detected in eight patients (15.4%) with diarrhea by the MRT-PCR and MPCR methods. The results of this study showed that four out of eight strains (50%) were STI a producer, and three out of eight strains were ST producers (37.5%). Also, one strain (12.5%) contained both STΙa and LT genes, simultaneously.

Conclusions: This is the first study performed in Shiraz to identify ETEC intestinal pathogens in patients with diarrhea. The results of this study showed that MRT-PCR can be used as a replacement for the conventional MPCR assay to detect the ETEC strains.

Implication for health policy/practice/research/medical education:

The prevalence and epidemiological features of enterotoxigenic Escherichia coli as a causative agent of diarrhea has a regional variation. A similar pattern has alsobeen observed between and within countries from the same geographical area.

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