Volume 16, Issue 62 (5-2026)
NCMBJ 2026, 16(62): 23-35 |
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Hassanifard M, Olad G, Amani J. Identification of B subunit of Cholera Toxin Using ELISA Technique |. NCMBJ 2026; 16 (62) :23-35
URL: http://ncmbjpiau.ir/article-1-1812-en.html
URL: http://ncmbjpiau.ir/article-1-1812-en.html
Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
Abstract: (18 Views)
Background and Aim: Vibrio cholerae is a Gram-negative bacterium that causes cholera, a disease characterized by severe diarrhea and dehydration due to the production of cholera toxin. Rapid and accurate detection of this toxin is essential for effective disease control. This study presents an ELISA-based method that is rapid, accurate, and cost-effective for detecting V. cholerae. This method involves producing a recombinant cholera toxin B subunit (CtxB) protein and generating specific antibodies in mice and rabbits.
Materials and Methods: The recombinant CtxB gene was expressed in E. coli BL21 (DE3) using IPTG induction and confirmed by Western blot analysis. The purified rCtxB protein was then administered subcutaneously to rabbits and mice. The antibodies were then isolated and purified. Their sensitivity was assessed using a sandwich ELISA, and their specificity was evaluated using an indirect ELISA against similar toxins.
Results: In this study, we used the ELISA technique with antibodies extracted and purified from the sera of mice and rabbits. Following colorimetric analysis, the optical density results demonstrated that the produced antibodies could specifically recognize the CtxB toxin. The method's high sensitivity (detecting eight nanograms of toxin) and minimal cross-reactivity with other toxins (StxB and LtB) confirm the antibodies' suitable specificity.
Conclusion: The protein- and antibody-based ELISA technique offers a rapid, accurate, and cost-effective approach for detecting V. cholerae compared to microbial culture methods or advanced molecular techniques. This method enables rapid detection of cholera toxin in clinical and environmental samples due to its high sensitivity, suitable specificity, and lack of requirement for advanced laboratory equipment.
Materials and Methods: The recombinant CtxB gene was expressed in E. coli BL21 (DE3) using IPTG induction and confirmed by Western blot analysis. The purified rCtxB protein was then administered subcutaneously to rabbits and mice. The antibodies were then isolated and purified. Their sensitivity was assessed using a sandwich ELISA, and their specificity was evaluated using an indirect ELISA against similar toxins.
Results: In this study, we used the ELISA technique with antibodies extracted and purified from the sera of mice and rabbits. Following colorimetric analysis, the optical density results demonstrated that the produced antibodies could specifically recognize the CtxB toxin. The method's high sensitivity (detecting eight nanograms of toxin) and minimal cross-reactivity with other toxins (StxB and LtB) confirm the antibodies' suitable specificity.
Conclusion: The protein- and antibody-based ELISA technique offers a rapid, accurate, and cost-effective approach for detecting V. cholerae compared to microbial culture methods or advanced molecular techniques. This method enables rapid detection of cholera toxin in clinical and environmental samples due to its high sensitivity, suitable specificity, and lack of requirement for advanced laboratory equipment.
Type of Study: Research Article |
Subject:
Genetics
Received: 2026/06/8 | Accepted: 2026/05/31 | Published: 2026/05/31
Received: 2026/06/8 | Accepted: 2026/05/31 | Published: 2026/05/31
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