A Biomarker Based Peptide Immunoassay for Clostridioides difficile: “Insights from Central India”

Biswas, Rima; Dhok, Archana; Nayak, Amit; Monaghan, Tanya M.; Kashyap, Rajpal Singh

Abstract

Introduction
Clostridioides difficile infection (CDI) is a major healthcare challenge due to its virulence factors, Toxins A and B. Current diagnostic methods like NAAT and EIA face limitations, including overdiagnosis and cultural resistance to stool sample collection, particularly in India. This study explores blood-based diagnostics, focusing on detecting anti-toxin antibodies through advanced proteomics and immunoassays. These innovative approaches aim to improve diagnostic sensitivity, specificity, and patient accessibility, addressing both clinical and cultural barriers.

Methods
This prospective observational study was conducted at the Advanced Research Centre of the Central India Institute of Medical Sciences (CIIMS) in Nagpur. The study enrolled 350 patients aged 18–70 years with clinical manifestations of diarrhea. This research focused on methodologies including microbial isolation of Clostridiodes difficile, isolating and analyzing novel proteins through LC-MS/MS, designing and synthesizing antigenic peptides, and standardizing peptide ELISA.

Results
The study successfully isolated and analyzed toxins A and B from C. difficile. The toxins were visualized using a 10% SDS-PAGE gel matrix, followed by peptide design and analysis. The developed immunoassay was tested on 350 serum samples, revealing a higher prevalence of toxin A than toxin B in the central Indian population.

Conclusions
The peptide-based immunoassay developed in this study marks a notable improvement in diagnosing Clostridioides difficile infection, especially in contexts where stool sample testing is impractical or culturally sensitive. Offering rapid, sensitive, and patient-friendly detection of anti-toxin antibodies, this method shows potential for enhancing CDI management and controlling its spread. However, additional refinement and validation are necessary to confirm its standalone diagnostic utility. The findings also underscore the intricate relationship between bacterial virulence, host immunity, and clinical outcomes, opening avenues for personalized treatments.

Citation

Biswas, R., Dhok, A., Nayak, A., Monaghan, T. M., & Kashyap, R. S. (2025). A Biomarker Based Peptide Immunoassay for Clostridioides difficile: “Insights from Central India”. International Journal of Peptide Research and Therapeutics, 31(3), Article 50. https://doi.org/10.1007/s10989-025-10709-y

• Journal Article Type: Article
• Acceptance Date: Feb 23, 2025
• Online Publication Date: Mar 12, 2025
• Publication Date: Mar 12, 2025
• Deposit Date: Mar 15, 2025
• Publicly Available Date: Mar 25, 2025
• Journal: International Journal of Peptide Research and Therapeutics
• Print ISSN: 1573-3149
• Electronic ISSN: 1573-3904
• Publisher: Springer Verlag
• Peer Reviewed: Peer Reviewed
• Volume: 31
• Issue: 3
• Article Number: 50
• DOI: https://doi.org/10.1007/s10989-025-10709-y
• Public URL: https://nottingham-repository.worktribe.com/output/46573567
• Publisher URL: https://link.springer.com/article/10.1007/s10989-025-10709-y
• Additional Information: Accepted: 23 February 2025; First Online: 12 March 2025; : ; : The authors declare no competing interests.