Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis

Negm, Ola H. and Hamed, Mohamed R. and Dilnot, Elizabeth M. and Shone, Clifford C. and Marszalowska, Izabela and Lynch, Mark and Loscher, Christine E. and Edwards, Laura J. and Tighe, Patrick J. and Wilcox, Mark H. and Monaghan, Tanya M. (2015) Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis. Clinical and Vaccine Immunology, 22 (9). pp. 1033-1039. ISSN 1556-679X

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Abstract

Clostridium difficile is an anaerobic, Gram-positive, and spore-forming bacterium that is the leading worldwide infective cause of hospital-acquired and antibiotic-associated diarrhea. Several studies have reported associations between humoral immunity and the clinical course of C. difficile infection (CDI). Host humoral immune responses are determined using conventional enzyme-linked immunosorbent assay (ELISA) techniques. Herein, we report the first use of a novel protein microarray assay to determine systemic IgG antibody responses against a panel of highly purified C. difficile-specific antigens, including native toxins A and B (TcdA and TcdB, respectively), recombinant fragments of toxins A and B (TxA4 and TxB4, respectively), ribotypespecific surface layer proteins (SLPs; 001, 002, 027), and control proteins (tetanus toxoid and Candida albicans). Microarrays were probed with sera from a total of 327 individuals with CDI, cystic fibrosis without diarrhea, and healthy controls. For all antigens, precision profiles demonstrated<10% coefficient of variation (CV). Significant correlation was observed between microarray and ELISA in the quantification of antitoxin A and antitoxin B IgG. These results indicate that microarray is a suitable assay for defining humoral immune responses to C. difficile protein antigens and may have potential advantages in throughput, convenience, and cost.

Item Type: Article
Keywords: Clostridium difficile, Antigen Microarray
Schools/Departments: University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Medicine > Nottingham Digestive Diseases Centre
University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Medicine
University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Life Sciences
Identification Number: https://doi.org/10.1128/CVI.00190-15
Depositing User: Negm, Ola
Date Deposited: 07 Dec 2015 15:10
Last Modified: 14 Sep 2016 06:56
URI: http://eprints.nottingham.ac.uk/id/eprint/31003

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