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:: Volume 10, Number 10 (December 2017) ::
Qom Univ Med Sci J 2017, 10(10): 25-33 Back to browse issues page
Design and Optimization of PCR Assay for Rapid Detection of Enterotoxigenic and Enteroaggregative Pathotypes of E. coli
Mohammad Soleimani1, Abbas Morovvati *2, Ali Javadi3
1- Qom Branch, Islamic Azad University
2- Qom Branch, Islamic Azad University , abbasmorovvati@gmail.com
3- Tehran University of Medical Sciences
Abstract:   (482 Views)

Background and Objectives: Escherichia coli is one of the most important causative agents of acute diarrheal disease in the world. Diarrheal diseases are one of the main causes of mortality among children in developing countries. The aim of this study was to develop a PCR technique for diagnosis of specific virulence genes of Enterotoxigenic E. coli (ETEC) and Enteroaggregative E. coli (EAEC).

Methods: In this study, specific genes ST and LT were used for diagnosis of ETEC pathotype and AAF/II specific gene was used for diagnosis of EAEC pathotype. The PCR technique was designed and optimized for these three genes. The reaction products were cloned in pTZ57R/T vector as positive control. The primers’ sensitivity was determined by measurement of detection limit, and primers’ specificity was evaluated using other bacterial genomes.

Results: According to the results of the PCR products analysis using agarose gel electrophoresis, the bands for AAF/II, LT, and ST were obtained 384, 459, and 150 bp, respectively. Also, the reaction of the genomes of control bacteria using the designed primers was negative. The detection limit based on copy number for ST, LT, and genes, was obtained 390, 1500, and 2500 copies, respectively.

Conclusion: Considering the findings of this study, design of these assays in Iran is a progress in the improving rapid and accurate diagnosis of pathogens and detection of E. coli pathotypes.

Keywords: Escherichia coli, Polymerase chain reaction, Dysentery
Full-Text [PDF 637 kb]   (339 Downloads)    
Type of Study: Original Article | Subject: میکروب شناسی
Received: 2016/02/24 | Accepted: 2016/04/17 | Published: 2016/12/25
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Soleimani M, Morovvati A, Javadi A. Design and Optimization of PCR Assay for Rapid Detection of Enterotoxigenic and Enteroaggregative Pathotypes of E. coli . Qom Univ Med Sci J . 2017; 10 (10) :25-33
URL: http://journal.muq.ac.ir/article-1-330-en.html
Volume 10, Number 10 (December 2017) Back to browse issues page
مجله دانشگاه علوم پزشکی قم Qom University of Medical Sciences Journal
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