The main objective of this work was to develop an antigen-capture enzyme-linked immunosorbent assay for detection of beta2-toxin in the intestine of neonatal piglets. The format of the assay comprised of capture antibodies (polyclonal), antigen (beta2-toxin), detecting antibody (labeled monoclonal) and a substrate. The ELISA was optimized using recombinant protein. After intestinal content samples were applied, the test protocol needed to be adjusted because of the presence of high background signal in some samples consistent with intestinal proteases. This was overcome by processing the samples at 4oC and using citrate buffer pH 6.1 containing 5% bovine serum albumin.
The second objective was to identify cpb2 in Clostridium perfringens type A isolates from selected animal species and to examine genotype-phenotype corelation. The study concluded that consensus cpb2, if present, was almost always expressed. In contrast, only about three-quarters of atypical cpb2, mostly was present in isolates of non-porcine origin, were expressed. / Ontario Ministry of Agriculture, Food and Rural Affairs; The Natural Sciences and Engineering Research Council of Canada
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OGU.10214/3495 |
| Date | 13 April 2012 |
| Creators | Kircanski, Jasmina |
| Contributors | Prescott, John |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Thesis |
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