Molecular epidemiology of salmonella typhi in Hong Kong.

by Norman Wai-sing Lo.Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.Includes bibliographical references (leaves 145-160).Chapter Chapter 1 --- Introduction --- p.1Chapter A. --- Classification of Salmonella --- p.3Chapter B. --- Enteric fever --- p.4Chapter C. --- Chloramphenicol and multiple resistance in S. Typhi --- p.7Chapter D. --- Resistance plasmids in the typhoid bacillus --- p.11Chapter E. --- In vivo acquisition of resistance plasmids by S. Typhi --- p.18Chapter F. --- Worldwide distribution of typhoid fever --- p.23Chapter G. --- Epidemiology of typhoid fever in Hong Kong --- p.27Chapter H. --- Principles of methods used in the epidemiological typing of S. Typhi --- p.29Chapter 1. --- Phage typing --- p.29Chapter 2. --- Antibiotics sensitivity pattern --- p.32Chapter 3. --- Plasmid analyses --- p.32Chapter a. --- Genetic methods --- p.33Chapter i. --- Phenotypic expression --- p.33Chapter ii. --- Transferability of plasmids --- p.33Chapter iii. --- Incompatibility --- p.34Chapter b. --- Molecular methods --- p.34Chapter i. --- Estimation of molecular size by agarose gel electrophoresis --- p.34Chapter ii. --- Plasmid fingerprinting --- p.35Chapter iii. --- Localization of resistance genes by DNA-DNA hybridization --- p.35Chapter 4. --- Ribotyping --- p.36Chapter 5. --- Total DNA fingerprinting --- p.37Chapter 6. --- Chromosomal insertion sequence IS200 profile analysis --- p.38Chapter 7. --- Others --- p.39Chapter a. --- Multilocus enzyme electrophoresis --- p.39Chapter b. --- Phenotype of lipopolysaccharide (LPS) --- p.39Chapter c. --- Envelope protein profiles and immunoblotting --- p.40Chapter I. --- Epidemiological typing of S. Typhi --- p.40Chapter J. --- Objectives --- p.46Chapter Chapter 2 --- Materials and Methods --- p.48Chapter A. --- Materials --- p.48Chapter I. --- Bacterial strains --- p.48Chapter II. --- Materials --- p.51Chapter B. --- Methods --- p.56Chapter I. --- Sensitivity testing --- p.56Chapter II. --- Characterization of plasmids --- p.58Chapter 1. --- Genetic studies --- p.58Chapter a. --- Transferability of resistance plasmids --- p.58Chapter b. --- Mobilization of resistances --- p.59Chapter c. --- Incompatibility grouping --- p.59Chapter 2. --- Molecular studies --- p.60Chapter a. --- Plasmid profile analysis --- p.60Chapter i. --- Plasmid extraction --- p.60Chapter ii. --- Agarose gel electrophoresis --- p.61Chapter b. --- Molecular characterization of plasmid --- p.61Chapter i. --- Extraction of purified plasmid DNA --- p.62Chapter ii. --- Restriction endonuclease digestion of plasmid DNA --- p.63Chapter III. --- Characterization β-lactamases --- p.63Chapter 1. --- Extraction of β-lactamases --- p.63Chapter 2. --- Determination of isoelectric points (pIs) --- p.64Chapter IV. --- Localization of resistance genes --- p.64Chapter 1. --- TEM-1 and TEM-2 genes --- p.65Chapter a. --- Oligonucleotide probes --- p.65Chapter b. --- Total DNA preparation --- p.66Chapter c. --- DNA-DNA-hybridization --- p.66Chapter i. --- Transfer of DNA to membrane filters --- p.66Chapter ii. --- Labelling of oligonucleotide probes (TEM-1 and TEM-2) --- p.67Chapter iii. --- Hybridization --- p.68Chapter 2. --- Chloramphenicol- and tetracycline-resistance genes --- p.69Chapter a. --- Preparation of probes --- p.69Chapter b. --- Hybridization --- p.73Chapter i. --- Transfer of DNA to membrane filters --- p.73Chapter ii. --- Labelling of chloramphenicol- and tetracycline- resistance probes --- p.73Chapter V. --- Epidemiological typing --- p.75Chapter 1. --- Ribotyping --- p.75Chapter 2. --- DNA fingerprinting --- p.77Chapter C. --- Plan for achieving objectives --- p.77Chapter Chapter 3 --- Results --- p.79Chapter A. --- Antimicrobial susceptibilities --- p.79Chapter B. --- Characterization of resistance plasmids --- p.83Chapter C. --- β-lactamases produced by ampicillin-resistant S. Typhi --- p.85Chapter D. --- Localization of resistance genes --- p.85Chapter E. --- Plasmid profile analysis --- p.87Chapter F. --- Plasmid fingerprinting --- p.90Chapter G. --- Ribotyping --- p.93Chapter H. --- Chromosomal DNA fingerprinting --- p.107Chapter I. --- "Correlation of PstI, ClaI and KpnI ribotypes and NarI, EcoRV and MluI chromosomal types" --- p.124Chapter J. --- Correlation of PstI ribotypes and NarI chromosomal types --- p.126Chapter K. --- Epidemiology of S. Typhi in Hong Kong --- p.129Chapter Chapter 4 --- Discussion --- p.131Chapter A. --- Antimicrobial susceptibilities --- p.131Chapter B. --- Characterization of resistances --- p.132Chapter C. --- Plasmid profile analysis --- p.135Chapter D. --- Epidemiological analysis of S. Typhi --- p.136Chapter E. --- Area for future research --- p.143References --- p.145Appendix --- p.161Chapter A. --- Buffer and Stock solutions --- p.161Chapter B. --- Epidemiological information of S. Typhi isolates --- p.16