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Title: Cystic fibrosis microbiology : molecular fingerprinting of microbial pathogens
Author: Doherty, C. J.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2000
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CF lung infections are caused by a surprisingly narrow spectrum of pathogens and include Staphylococcus aureus, non-capsulate Haemophilus influenzae, Pseudomonas aeruginosa and Burkholderia cepacia. Stenotrophomonas maltophila is recovered from respiratory secretions with increasing frequency; however, its pathogenic role remains unclear. The primary aims of this thesis include the development and use of genomic fingerprinting systems to assist epidemiological investigations of CF pathogens, including S. maltophilia. Genomic fingerprinting is based on digestion of total bacterial chromosomal DNA with rare cutting enzymes, chosen on the basis of the bacterium's GC content. Separation of the DNA fragments, is then achieved by pulsed-field gel electrophoresis (PFGE) in an appropriate apparatus such as the Bio-Rad contour clamped homogeneous electric field (CHEF) system. Although a variety of other genomic typing systems are available, the thesis focused on PFGE, potentially the most discriminating system at present. Another major theme of the thesis concerned the epidemiology of B. cepacia. This highly adaptable plant and human pathogen causes great anxiety in the CF community on account of its inherent resistance, transmissibility and association with cepacia syndrome, a rapidly fatal pneumonia affecting approximately 30% of colonised patients. PFGE is technically demanding, time consuming and relatively expensive, thus attempts were made to assess the reliability and potential of other systems, in particular, PCR-ribotyping as a simple and rapid screening system for clonal analyses. The project provided a limited opportunity for fingerprinting and other microbiological studies of the commensal and pathogenic respiratory flora in CF patients participating in the first human trials of CF gene therapy. Specimens were examined before, during and after local nasal administration of a DNA/liposome complex. Although only a Phase 1 study was achieved during the duration of the thesis, microbiological analyses provided interesting results, in particular an unexpected lack of clonal relationship between S. aurens colonising the upper and lower airways.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available