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Introduction <p><i>Clostridioides difficile</i> 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.</p> Methods <p>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 <i>Clostridiodes difficile</i>, isolating and analyzing novel proteins through LC-MS/MS, designing and synthesizing antigenic peptides, and standardizing peptide ELISA.</p> Results <p>The study successfully isolated and analyzed toxins A and B from <i>C. difficile</i>. 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.</p> Conclusions <p>The peptide-based immunoassay developed in this study marks a notable improvement in diagnosing <i>Clostridioides difficile</i> 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.</p>

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

  • 期刊论文

  • 发表日期：

    2025年3月12日

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

摘要

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.