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A Protein Microarray Assay for Serological Determination of Antigen-specific Antibody Responses Following Clostridium difficile Infection
Published on: June 15, 2018
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Profiling Humoral Immune Responses to Clostridium difficile-Specific Antigens by Protein Microarray Analysis
Ola H Negm1, Mohamed R Hamed1, Elizabeth M Dilnot2
1Immunology, School of Life Sciences, University of Nottingham, Nottingham, United Kingdom Medical Microbiology and Immunology, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
Clinical and Vaccine Immunology : CVI
|July 17, 2015
Summary
A novel protein microarray assay effectively measures IgG antibody responses to Clostridium difficile antigens. This method shows promise for improved throughput and cost-efficiency in diagnosing C. difficile infection (CDI).
Area of Science:
- Immunology
- Microbiology
- Biotechnology
Background:
- Clostridium difficile is a major cause of hospital-acquired and antibiotic-associated diarrhea.
- Humoral immunity plays a role in the clinical course of C. difficile infection (CDI).
- Conventional methods like ELISA are used to assess host immune responses.
Purpose of the Study:
- To introduce and validate a novel protein microarray assay for detecting systemic IgG antibody responses to C. difficile antigens.
- To compare the performance of the microarray assay with traditional ELISA techniques.
Main Methods:
- A protein microarray was developed using purified C. difficile antigens (toxins A and B, fragments, surface layer proteins) and control proteins.
- The microarray was probed with sera from 327 individuals, including those with CDI, cystic fibrosis, and healthy controls.
- Precision was assessed by coefficient of variation (<10%), and microarray results were correlated with ELISA for antitoxin A and B IgG.
Main Results:
- The protein microarray demonstrated high precision (<10% CV) for all tested C. difficile antigens.
- Significant correlation was found between the microarray and ELISA for quantifying IgG antibodies against C. difficile toxins A and B.
- The assay successfully detected systemic IgG antibody responses in individuals with CDI.
Conclusions:
- The novel protein microarray assay is a suitable method for defining humoral immune responses to C. difficile protein antigens.
- This microarray technology offers potential advantages in terms of throughput, convenience, and cost compared to conventional assays.
- The assay may be valuable for research and clinical diagnostics related to C. difficile infection.
