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Title: Genotypic diversity and epidemiological typing of Bordetella pertussis
Author: Neal, Shona Elaine
ISNI:       0000 0001 3440 4116
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 2004
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The resurgence of pertussis in several highly vaccinated countries has stimulated this study of the genotypic diversity and epidemiological typing of Bordetella pertussis. The genotypic variation of B. pertussis was investigated in 318 UK clinical isolates from 1920-2002, vising pertactin (prnA) and pertussis toxin SI subunit (ptxA) gene typing. To date, the evaluation of typing methods used for B. pertussis has not been performed extensively. Therefore, in this thesis, the recognised methods, namely serotyping, pulsed-field gel electrophoresis (PFGE) using Xbal, and IS1002-RFLP analysis, were evaluated. It was found that, PFGE typing gave a good discrimination index value, but gave a low score for reproducibility, and further work is required to optimise this method. Furthermore, if prnA and ptxA gene typing were included in the evaluation, combined with serotyping, this combination would equal the discrimination of PFGE. Other typing methods attempted for B. pertussis included direct sequencing of adenylate kinase (adk) and filamentous haemagglutinin genes (fhaB), and single-enzyme amplified fragment length polymorphism (AFLP) analysis with a selection of enzymes and selective primers. The type strain and a clinical strain, generated one and six single nucleotide polymorphisms (SNPs) in adk and fhoB, respectively. The discriminatory ability of the single-enzyme AFLP analysis was not satisfactory, as only a few different profiles were seen in the nine isolates tested. However, at least four AFLP profiles were generated using PstI enzyme, and the selective primer Pst-C. The direct detection and epidemiological typing of B. pertussis in 20 previously untypable clinical samples was attempted using prnA, ptxA as targets. Six clinical extracts generated prnA and ptxA (5/6) sequence data, therefore confirming that these typing procedures on B. pertussis PCR-positive clinical specimens is worthwhile in order to generate the prnA and ptxA distributions from babies or adults presenting atypical symptoms. This strategy should also be encouraged in other country that have studied prnA and ptxA allele distributions, in order to update the representation of the genetic diversity of B. pertussis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available