Abbas Hajizade *(1), Peyman - PowerPoint Presentation

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Abbas Hajizade*(1), Peyman Avakh (1), Shahram Nazarian (1), Yousof Tarvrdi-zadeh (1)1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.*Corresponding Author. E-mail: abbashajizade@gmail.com

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Abbas

Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.

Simultaneous detection and discrimination of enteropathogenic E. coli using multiplex-PCR

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V_Poster Code: A-10-408-2

Introduction13/OCT/10

2Abbas Hajizade

- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.

Enteropathogenic E. coli are diarrhagenic bacteria responsible for many diarrheal cases all over the world. Indeed, some of them have the ability to be exploited as bioterrorism agent. There are six pathotypes of diarrhagenic E. coli : enterotoxigenic E. coli , enterohemorrhagic E. coli , enterntroinvasive

E. coli, enteropathogenic E. coli

, enteroaggregative E. coli , and diffusely adherent E. coli . Though enterohemorrhagic E. coli and enterotoxigenic E. coli are the most potent pathotypes which are considered as the potential disabilitating microbial agents, however, other pathotypes are also important in this regard. Water and food resources can be contaminated by these bacteria, so detection of these bacteria can protect the personnel against a bioterrorism disaster. In the present study a multiplex-PCR was developed for simultaneous detection and discrimination of different enteropathogenic E. coli

pathotypes.

Method13/OCT/10

3Abbas Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.

For this aim, firstly the specific gene(s) of each

pathotype was selected and retrieved from

GeneBank. Then, appropriate primers were designed for amplification of a special region of each gene. The primers were designed so that the resultant fragment of each gene had a defined length and the fragments' lengths were unique. Indeed, primers were designed to be compatible to be used in a multiplex PCR reaction. At last, the primers were used for the detection and discrimination of enteropathogenic E. coli.

Result

The selected genes were as follow:lt and sta genes for ETEC, eae and bfp genes for EPEC,

stx of EHEC, pCVD and

aggR

genes for EAEC, virF and ipaH genes for EIEC, and daad gene for DAEC.

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Abbas Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.

Conclusion13/OCT/10

5Abbas Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.

The result shows that the designed primers have the ability to detect the pathogenic E. coli

strains and it is possible to detect and discriminate

enteropathogenic E. coli in a single reaction tube.

References

Moyo SJ, Maselle SY, Matee MI,Langeland N,

Mylvaganam H.Identification of diarrheagenic

Escherichiacoli isolated from infants and children indares Salaam Tanzania. BMC Infect Dis2007;7:92-7. Nataro JP. Diarrhea among children in developing countries.AdvExp Med Biol 2013; 764:73-80. Mohammadzadeh M, Goudarzi H, Dabiri H, Fallah

F. Frequency of Enteroaggregative Escherichia coli isolates among patients with diarrhea referred to Tehran hospitals. Research in Medicine. 2016; 40 (2) :68-72. Nguyen, T. V., Le Van, P., Le Huy, C.,

Gia, K. N. & Weintraub, A. Detection and characterization of diarrheagenic Escherichia coli from young children in Hanoi, Vietnam. J Clin Microbiol. 2005: 43, 755– 760.O’Ryan M, Lucero Y, O’Ryan-Soriano MA, Ashkenazi S. An update on management of severe acute infectious gastroenteritis in children. Expert Rev Anti Infect Ther 2010; 8(6):671-82.13/OCT/10

6Abbas Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam

Hossein University, Tehran, Iran.

Final Questions

Using specific primers, it is possible to discriminate between different bacterial strains. There are some suggested primers to detect and discriminate enteropathogenic E. coli. In the present study a new set of primers were designed to detect and discriminate

enteropathogenic E. coli in one reaction tube. The resulted amplicons

have different lengths so that they are proper for this aim.

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Abbas Hajizade- 1Biology Research Center, Faculty of Basic Sciences, Imam Hossein University, Tehran, Iran.