Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis
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 infecti...
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| Format: | Article |
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American Society of Microbiology
2015
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| Online Access: | https://eprints.nottingham.ac.uk/31003/ |
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| author | Negm, Ola H. Hamed, Mohamed R. Dilnot, Elizabeth M. Shone, Clifford C. Marszalowska, Izabela Lynch, Mark Loscher, Christine E. Edwards, Laura J. Tighe, Patrick J. Wilcox, Mark H. Monaghan, Tanya M. |
| author_facet | Negm, Ola H. Hamed, Mohamed R. Dilnot, Elizabeth M. Shone, Clifford C. Marszalowska, Izabela Lynch, Mark Loscher, Christine E. Edwards, Laura J. Tighe, Patrick J. Wilcox, Mark H. Monaghan, Tanya M. |
| author_sort | Negm, Ola H. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:10:57Z |
| format | Article |
| id | nottingham-31003 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:10:57Z |
| publishDate | 2015 |
| publisher | American Society of Microbiology |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-310032024-08-15T15:32:34Z https://eprints.nottingham.ac.uk/31003/ Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis Negm, Ola H. Hamed, Mohamed R. Dilnot, Elizabeth M. Shone, Clifford C. Marszalowska, Izabela Lynch, Mark Loscher, Christine E. Edwards, Laura J. Tighe, Patrick J. Wilcox, Mark H. Monaghan, Tanya M. 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. American Society of Microbiology 2015-09 Article PeerReviewed Negm, Ola H., Hamed, Mohamed R., Dilnot, Elizabeth M., Shone, Clifford C., Marszalowska, Izabela, Lynch, Mark, Loscher, Christine E., Edwards, Laura J., Tighe, Patrick J., 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 Clostridium difficile Antigen Microarray http://cvi.asm.org/content/22/9/1033.full doi:10.1128/CVI.00190-15 doi:10.1128/CVI.00190-15 |
| spellingShingle | Clostridium difficile Antigen Microarray Negm, Ola H. Hamed, Mohamed R. Dilnot, Elizabeth M. Shone, Clifford C. Marszalowska, Izabela Lynch, Mark Loscher, Christine E. Edwards, Laura J. Tighe, Patrick J. Wilcox, Mark H. Monaghan, Tanya M. Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title | Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title_full | Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title_fullStr | Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title_full_unstemmed | Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title_short | Profiling humoral immune responses to Clostridium difficile-specific antigens by protein microarray analysis |
| title_sort | profiling humoral immune responses to clostridium difficile-specific antigens by protein microarray analysis |
| topic | Clostridium difficile Antigen Microarray |
| url | https://eprints.nottingham.ac.uk/31003/ https://eprints.nottingham.ac.uk/31003/ https://eprints.nottingham.ac.uk/31003/ |