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:: Volume 15, Number 1 (April-May 2012) ::
amuj 2012, 15(1): 35-42 Back to browse issues page
Production and purification of heat-labile toxin of enterotoxigenic Escherichia coli and its detection by GM1 gangelioside receptor-ELISA based method
Raziyeh Khalesi, Jafar Salimian , Shahram Nazarian, Zahra Ehsaei, Ali asghar Rahimi, Nafiseh Amini, Seyed Mohammad Moazzeni
PhD , jafarsalimyan@yahoo.com
Abstract:   (8531 Views)
Background: Enterotoxigenic Escherichia coli bacterium is the most important bacterial agent causing diarrhea. Specific virulence factors, such as enterotoxins and colonization factors, distinguish ETEC from other classes of diarrheagenic E.coli. In this study, heat-labile toxin was purified which could be utilized for anti-toxin assay in GM1 gangelioside receptor-ELISA based method and for identification of ETEC producing toxin. Materials and Methods: In this experimental study, bacterial strain producing heat-labile toxin was first cultivated for production and purification of toxin. Then supernatant soluble proteins were precipitated with ammonium sulfate and purified using biochemical methods. Finally, purified protein was dialyzed against Tris 0.02 mM pH 8 and analyzed on gel electrophoresis. GM1 gangelioside receptor-ELISA based method was used for detection and assessment of the purified toxin. Through this method, the effect of anti-recombinant heat-labile toxin B subunit neutralization on heat-labile toxin was investigated. Results: Toxin purification was revealed by the presence of 12 and 28 KD protein bands. This study demonstrated that anti-recombinant heat-labile toxin B subunit antibody can detect the purified toxin and can inhibit its binding to GM1 receptor up to 80%. Conclusion: Purification of heat-labile toxin and gangelioside receptor-ELISA assay can be used for accurate detection and epidemiological study of clinical isolates.
Keywords: Antibody against recombinant B subunit, Enterotoxigenic Escherichia coli, GM1 gangelioside receptor-ELISA, Heat-labile enterotoxin
Full-Text [PDF 367 kb]   (2151 Downloads)    
Type of Study: Original Atricle | Subject: Basic Sciences
Received: 2011/01/15
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Khalesi R, Salimian J, Nazarian S, Ehsaei Z, Rahimi A A, Amini N et al . Production and purification of heat-labile toxin of enterotoxigenic Escherichia coli and its detection by GM1 gangelioside receptor-ELISA based method. amuj. 2012; 15 (1) :35-42
URL: http://amuj.arakmu.ac.ir/article-1-1010-en.html
Volume 15, Number 1 (April-May 2012) Back to browse issues page
مجله دانشگاه علوم پزشکی اراک Arak Medical University Journal
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