PRODUCTION AND PURIFICATION OF HEAT-LABILE TOXIN OF ENTEROTOXIGENIC ESCHERICHIA COLI AND ITS DETECTION BY GM1 GANGELIOSIDE RECEPTOR-ELISA BASED METHOD

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KHALESI R.; SALIMIAN J.; NAZARIAN SH.; EHSAEI Z.; RAHIMI A.A.; AMINI N.; MOAZZENI S.M.

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Journal: Journal of Arak University Medical Sciences Year:2012 | Volume:15 | Issue:1 (60) Page(s): 35-42

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Title

PRODUCTION AND PURIFICATION OF HEAT-LABILE TOXIN OF ENTEROTOXIGENIC ESCHERICHIA COLI AND ITS DETECTION BY GM1 GANGELIOSIDE RECEPTOR-ELISA BASED METHOD

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Keywords

ANTIBODY AGAINST RECOMBINANT B SUBUNITQ2

ENTEROTOXIGENIC ESCHERICHIA COLIQ2

GM1 GANGELIOSIDE RECEPTOR-ELISAQ2

HEAT-LABILE ENTEROTOXINQ2

Abstract

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.

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APA: Copy

KHALESI, R., SALIMIAN, J., NAZARIAN, SH., EHSAEI, Z., RAHIMI, A.A., AMINI, N., & MOAZZENI, S.M.. (2012). PRODUCTION AND PURIFICATION OF HEAT-LABILE TOXIN OF ENTEROTOXIGENIC ESCHERICHIA COLI AND ITS DETECTION BY GM1 GANGELIOSIDE RECEPTOR-ELISA BASED METHOD. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), 15(1 (60)), 35-42. SID. https://sid.ir/paper/68944/en

Vancouver: Copy

KHALESI R., SALIMIAN J., NAZARIAN SH., EHSAEI Z., RAHIMI A.A., AMINI N., MOAZZENI S.M.. PRODUCTION AND PURIFICATION OF HEAT-LABILE TOXIN OF ENTEROTOXIGENIC ESCHERICHIA COLI AND ITS DETECTION BY GM1 GANGELIOSIDE RECEPTOR-ELISA BASED METHOD. ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ)[Internet]. 2012;15(1 (60)):35-42. Available from: https://sid.ir/paper/68944/en

IEEE: Copy

R. KHALESI, J. SALIMIAN, SH. NAZARIAN, Z. EHSAEI, A.A. RAHIMI, N. AMINI, and S.M. MOAZZENI, “PRODUCTION AND PURIFICATION OF HEAT-LABILE TOXIN OF ENTEROTOXIGENIC ESCHERICHIA COLI AND ITS DETECTION BY GM1 GANGELIOSIDE RECEPTOR-ELISA BASED METHOD,” ARAK MEDICAL UNIVERSITY JOURNAL (AMUJ), vol. 15, no. 1 (60), pp. 35–42, 2012, [Online]. Available: https://sid.ir/paper/68944/en

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