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:: Volume 9, Issue 2 (summer2011 2011) ::
jmj 2011, 9(2): 1-7 Back to browse issues page
Pulsed field gel electrophoresis protocol for typing of uropathogenic Escherichia coli
Mojtaba Anvarinejad *, Shohreh Farshad, Vahhab Alborzi, Reza Ranjbar, Marzeyeh Hoseini
Abstract:   (13536 Views)

Introduction: Pulsed Field Gel Electrophoresis (PFGE) is one of the best typing methods which is highly effective in molecular epidemiological studies in different areas. All bacterial isolates are typeable by PFGE and the results are highly reproducible. As this technique is very sensitive and the least variation in the parameter leads to lack of result or false positive, in this study we tried to optimize this method and eliminate the technical problems to obtain accurate results for comparison of different strains in epidemiological studies. Material and Methods: To assess the genotype pattern of Escherichia coli strains by PFGE method, we studied different parameters such as concentration of DNA molecules of the sample and their relationship with the enzymes concentration. Results: After different examinations, the best optical density for bacterial suspension in the wavelength of 610 was found to be within 0.6-0.7 nm and the best concentration of XbaI enzyme was 150 units. The appropriate time for proteinase K effect was 7 hours. Conclusion: It seems that the parameters used in this study are the most essential factors in the establishment of an effective PFGE method. With due attention to this experiment, researchers can use this method more efficiently than before for epidemiologic studies and comparison of different strains.

Keywords: Escherichia coli, Electrophoresis, Gel, Pulsed Field
Full-Text [PDF 312 kb]   (4376 Downloads)    
Type of Study: Research |
Received: 2013/08/19 | Revised: 2018/03/3 | Accepted: 2013/08/21
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Anvarinejad M, Farshad S, Alborzi V, Ranjbar R, Hoseini M. Pulsed field gel electrophoresis protocol for typing of uropathogenic Escherichia coli. jmj. 2011; 9 (2) :1-7
URL: http://jmj.jums.ac.ir/article-1-176-en.html

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Volume 9, Issue 2 (summer2011 2011) Back to browse issues page
فصلنامه دانشگاه علوم پزشکی جهرم Journal of Jahrom University of Medical Sciences
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