Pectobacterium carotovorum ssp. carotovorum (Pcc) is implicated in the destruction of
ornamental plants in greenhouse recirculating systems. PCR-based detection and
quantification of Pcc requires expensive instrumentation and knowledgeable users. This
thesis describes the production of polyclonal antibodies and a single-domain antibody
fragment (VHH) against Pcc lipopolysaccharide (LPS), and the development of user-
friendly diagnostic assays for detection and quantification of the pathogen. Polyclonal
ELISAs against heat-killed (HK) Pcc (limit of detection (LOD) = 81 CFU/ml; limit of
quantitation (LOQ) = 216 CFU/ml) and Pcc LPS (LOD = 23 ng/ml; LOQ = 76 ng/ml)
were developed. A preliminary user-friendly dipstick assay was also developed (≥ 105
CFU/ml). A phage display library was constructed (6.0 x 105 clones/ml), yielding one
unique anti-Pcc LPS VHH. Using the Pcc LPS-specific VHH to produce affordable, user-
friendly diagnostic assays is feasible since antibody fragments can be produced on a large
scale through expression in Escherichia coli or Piccia pastoris. / Flowers Canada, CANADA-ONTARIO RESEARCH AND DEVELOPMENT (CORD) PROGRAM, Canada Research Chairs (CRC) Program, NSERC/NRC
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OGU.10214/2314 |
| Date | 28 October 2010 |
| Creators | Bassoriello, Melissa Maria Ivana |
| Contributors | Blom, Theo J., Hall, J. Christopher |
| 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 |
| Rights | http://creativecommons.org/licenses/by/3.0/ |
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