Volume 16, Issue 62 (5-2026)
NCMBJ 2026, 16(62): 23-35 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
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: (474 Views)
Extended Abstract
Background and Aim: Cholera is a major waterborne infectious disease caused by Vibrio cholerae, a Gram-negative bacterium that produces cholera toxin (CT), the main virulence factor responsible for severe diarrhea and dehydration. Rapid and accurate detection of this toxin is crucial for disease diagnosis, outbreak monitoring, and public health interventions. Conventional culture-based and molecular diagnostic methods are often expensive, time-consuming, and require specialized laboratory facilities.
Materials and Methods: Therefore, the development of a simple, sensitive, and cost-effective diagnostic method is highly desirable. This study aimed to develop an ELISA-based assay for cholera toxin detection using the recombinant cholera toxin B subunit (rCtxB) and specific polyclonal antibodies. The CtxB gene was expressed in Escherichia coli BL21(DE3) following induction with IPTG. Expression of the recombinant protein was confirmed by Western blot analysis. Purified rCtxB protein was used to immunize rabbits and mice via subcutaneous injections. Following immunization, serum samples were collected, and antibodies were purified.
Results:The sensitivity of the generated antibodies was evaluated using sandwich ELISA, while their specificity was assessed by indirect ELISA against structurally related toxins, including heat-labile enterotoxin B subunit (LtB) and Shiga toxin B subunit (StxB). The purified antibodies obtained from rabbits and mice demonstrated strong and specific reactivity toward CtxB. ELISA results confirmed successful recognition of cholera toxin with high sensitivity. The developed assay was able to detect as little as 8 ng of toxin. In addition, minimal cross-reactivity was observed with LtB and StxB, indicating excellent specificity. These findings demonstrate the effectiveness of the produced antibodies for cholera toxin detection.
Conclusion:The developed ELISA method provides a rapid, sensitive, accurate, and affordable approach for detecting cholera toxin. Compared with conventional microbiological and advanced molecular methods, this assay offers significant advantages in terms of simplicity, low cost, and ease of implementation. The method does not require sophisticated laboratory equipment and can be effectively applied for the detection of Vibrio cholerae toxin in clinical and environmental samples, supporting cholera surveillance and outbreak control programs.
Background and Aim: Cholera is a major waterborne infectious disease caused by Vibrio cholerae, a Gram-negative bacterium that produces cholera toxin (CT), the main virulence factor responsible for severe diarrhea and dehydration. Rapid and accurate detection of this toxin is crucial for disease diagnosis, outbreak monitoring, and public health interventions. Conventional culture-based and molecular diagnostic methods are often expensive, time-consuming, and require specialized laboratory facilities.
Materials and Methods: Therefore, the development of a simple, sensitive, and cost-effective diagnostic method is highly desirable. This study aimed to develop an ELISA-based assay for cholera toxin detection using the recombinant cholera toxin B subunit (rCtxB) and specific polyclonal antibodies. The CtxB gene was expressed in Escherichia coli BL21(DE3) following induction with IPTG. Expression of the recombinant protein was confirmed by Western blot analysis. Purified rCtxB protein was used to immunize rabbits and mice via subcutaneous injections. Following immunization, serum samples were collected, and antibodies were purified.
Results:The sensitivity of the generated antibodies was evaluated using sandwich ELISA, while their specificity was assessed by indirect ELISA against structurally related toxins, including heat-labile enterotoxin B subunit (LtB) and Shiga toxin B subunit (StxB). The purified antibodies obtained from rabbits and mice demonstrated strong and specific reactivity toward CtxB. ELISA results confirmed successful recognition of cholera toxin with high sensitivity. The developed assay was able to detect as little as 8 ng of toxin. In addition, minimal cross-reactivity was observed with LtB and StxB, indicating excellent specificity. These findings demonstrate the effectiveness of the produced antibodies for cholera toxin detection.
Conclusion:The developed ELISA method provides a rapid, sensitive, accurate, and affordable approach for detecting cholera toxin. Compared with conventional microbiological and advanced molecular methods, this assay offers significant advantages in terms of simplicity, low cost, and ease of implementation. The method does not require sophisticated laboratory equipment and can be effectively applied for the detection of Vibrio cholerae toxin in clinical and environmental samples, supporting cholera surveillance and outbreak control programs.
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
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
