Antibody-based detection and quantification of Pectobacterium carotovorum ssp. carotovorum

dc.contributor.advisorBlom, Theo J.
dc.contributor.advisorHall, J. Christopher
dc.contributor.authorBassoriello, Melissa Maria Ivana
dc.date.accessioned2010-10-28T17:07:38Z
dc.date.available2010-10-28T17:07:38Z
dc.date.copyright2010-10
dc.date.created2010-10-14
dc.date.issued2010-10-28T17:07:38Z
dc.degree.departmentDepartment of Environmental Biologyen_US
dc.degree.grantorUniversity of Guelphen_US
dc.degree.nameMaster of Scienceen_US
dc.degree.programmeEnvironmental Biologyen_US
dc.description.abstractPectobacterium 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_US
dc.description.sponsorshipFlowers Canada
dc.description.sponsorshipCANADA-ONTARIO RESEARCH AND DEVELOPMENT (CORD) PROGRAM
dc.description.sponsorshipCanada Research Chairs (CRC) Program
dc.description.sponsorshipNatural Sciences and Engineering Research Council of Canada/National Research Council
dc.identifier.urihttp://hdl.handle.net/10214/2314
dc.language.isoenen_US
dc.publisherUniversity of Guelphen_US
dc.rightsAttribution 3.0 Unported*
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/*
dc.subjectELISAen_US
dc.subjectantibodiesen_US
dc.subjectdetectionen_US
dc.subjectquantificationen_US
dc.subjectPectobacterium carotovorum ssp. carotovorumen_US
dc.titleAntibody-based detection and quantification of Pectobacterium carotovorum ssp. carotovorumen_US
dc.typeThesisen_US

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