ANTIBODY-BASED DETECTION AND QUANTIFICATION OF PECTOBACTERIUM CAROTOVORUM SSP. CAROTOVORUM

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ANTIBODY-BASED DETECTION AND QUANTIFICATION OF PECTOBACTERIUM CAROTOVORUM SSP. CAROTOVORUM

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dc.contributor.advisor Blom, Theo J.
dc.contributor.advisor Hall, J. Christopher
dc.contributor.author Bassoriello, Melissa Maria Ivana
dc.date 2010-10-14
dc.date.accessioned 2010-10-28T17:07:38Z
dc.date.available 2010-10-28T17:07:38Z
dc.date.issued 2010-10-28T17:07:38Z
dc.identifier.uri http://hdl.handle.net/10214/2314
dc.description.abstract 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. en
dc.description.sponsorship Flowers Canada, CANADA-ONTARIO RESEARCH AND DEVELOPMENT (CORD) PROGRAM, Canada Research Chairs (CRC) Program, NSERC/NRC en
dc.language.iso en en
dc.rights.uri http://creativecommons.org/licenses/by/3.0/ *
dc.subject ELISA, antibodies, detection, quantification, Pectobacterium carotovorum ssp. carotovorum en
dc.title ANTIBODY-BASED DETECTION AND QUANTIFICATION OF PECTOBACTERIUM CAROTOVORUM SSP. CAROTOVORUM en
dc.type Thesis en
dc.degree.programme Environmental Biology en
dc.degree.name Master of Science en
dc.degree.department Department of Environmental Biology en


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http://creativecommons.org/licenses/by/3.0/ Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/3.0/

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