
handle: 10214/2314 , 10214/19897
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.
Pectobacterium caratovorum ssp. caravorum, Pectobacterium carotovorum ssp. carotovorum, Recirculating systems, detection, antibodies, Polyclonal antibodies, ELISA, Greenhouses, quantification, Ornamental plants
Pectobacterium caratovorum ssp. caravorum, Pectobacterium carotovorum ssp. carotovorum, Recirculating systems, detection, antibodies, Polyclonal antibodies, ELISA, Greenhouses, quantification, Ornamental plants
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