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

Ola H. Negm, Mohamed R. Hamed, Elizabeth M. Dilnot, Clifford Shone, Izabela Marszalowska, Mark Lynch, Christine E. Loscher, Laura J. Edwards, Patrick J. Tighe, Mark H. Wilcox, Tanya M. Monaghan*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)


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 enzymelinked 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.

Original languageEnglish
Pages (from-to)1033-1039
Number of pages7
JournalClinical and Vaccine Immunology
Issue number9
Publication statusPublished - 1 Sep 2015


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