by Chu Ka-yi. / Thesis submitted in: October 2000. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 160-174). / Abstracts in English and Chinese. / ABSTRACT (English) --- p.i / ABSTRACT (Chinese) --- p.iii / ACKNOWLEDGMENT --- p.v / LIST OF CONTENTS --- p.vi / LIST OF TABLES --- p.x / LIST OF FIGURES --- p.xii / ABBREVIATIONS --- p.xiv / TERMS --- p.xv / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Taxonomy of Acinetobacter - historical and current --- p.1 / Chapter 1.2 --- Ecology and clinical significance of Acinetobacter --- p.5 / Chapter 1.3 --- General identification and typing methods for Acinetobacter species / Chapter 1.3.1 --- Identification at species level --- p.9 / Chapter 1.3.2 --- Identification at strain level --- p.11 / Chapter 1.4 --- Methods used in this study for characterization of Acinetobacter species / Chapter 1.4.1 --- Amplified ribosomal DNA restriction analysis (ARDRA) --- p.14 / Chapter 1.4.2 --- tDNA spacer fingerprinting (tDNA) --- p.15 / Chapter 1.4.3 --- Fluorescent amplified fragment length polymorphism (FAFLP) --- p.16 / Chapter 1.4.4 --- Phenotypic methods including carbon utilization tests --- p.20 / Chapter 1.5 --- Objectives --- p.25 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.27 / Chapter 2.1 --- Bacterial strains and isolates --- p.27 / Chapter 2.2 --- Materials / Chapter 2.2.1 --- Antimicrobial agents and chemicals --- p.30 / Chapter 2.2.2 --- "Carbohydrates, enzymes and other materials" --- p.32 / Chapter 2.2.3 --- Commercial media and media prepared manually --- p.33 / Chapter 2.2.4 --- "Buffers, solutions and list of instruments" --- p.35 / Chapter 2.3 --- General Bacteriological Techniques / Chapter 2.3.1 --- Isolation of acinetobacters --- p.38 / Chapter 2.3.2 --- Routine bacteriological identification --- p.39 / Chapter 2.4 --- General Molecular Biology Techniques / Chapter 2.4.1 --- DNA isolation --- p.40 / Chapter 2.4.2 --- Transformation --- p.41 / Chapter 2.4.3 --- Agarose gel electrophoresis --- p.43 / Chapter 2.5 --- Genospeciation of acinetobacters by Amplified Ribosomal Restriction DNA Analysis (ARDRA) --- p.44 / Chapter 2.6 --- Characterization of ARDRA unclassifiable acinetobacters (AUA) by Phenotypic methods / Chapter 2.6.1 --- Temperature tolerance tests --- p.47 / Chapter 2.6.2 --- Carbon utilization tests --- p.47 / Chapter 2.6.3 --- Gelatin and hemolysis tests --- p.48 / Chapter 2.6.4 --- Minimum Inhibitory Concentration (MIC) --- p.49 / Chapter 2.7 --- Characterization of AUA by tDNA spacer fingerprinting (tDNA) method --- p.51 / Chapter 2.8 --- Characterization of AUA by Fluorescent Amplified Fragment Length Polymorphism analysis (FAFLP) --- p.55 / Chapter 2.9 --- Relatedness study of isolates within the same AUA group by Enterobacterial Repetitive Intergenic Consensus (ERIC) typing method --- p.58 / Chapter CHAPTER 3 --- COLLECTION OF UNCLASSIFIABLE ACINETOBACTERS by ARDRA (AUA) METHOD --- p.59 / Chapter 3.1 --- Results / Chapter 3.1.1 --- Isolation and genospeciation of acinetobacters from hospital environments and raw food --- p.59 / Chapter 3.1.2 --- Collection of ARDRA unclassifiable acinetobacters (AUA) --- p.63 / Chapter 3.2 --- Discussion / Chapter 3.2.1 --- Limitations and merits of ARDRA method --- p.68 / Chapter 3.2.2 --- Potential significance of the representative AUA groups --- p.71 / Chapter CHAPTER 4 --- CHARACTERIZATION OF ARDRA UNCLASSIFIABLE ACINETOBACTERS (AUA) BY tDNA SPACER (tDNA) FINGERPRINTING METHOD --- p.72 / Chapter 4.1 --- Results / Chapter 4.1.1 --- Assessment of reproducibility --- p.72 / Chapter 4.1.2 --- Construction of the database with the reference strains --- p.75 / Chapter 4.1.3 --- Characterization of the representative AUA groups --- p.78 / Chapter 4.2 --- Discussion / Chapter 4.2.1 --- Evaluation of the reproducibility and discriminatory power --- p.89 / Chapter 4.2.2 --- Possible genospeciation of the representative AUA groups --- p.92 / Chapter 4.2.3 --- Limitations and merits --- p.96 / Chapter CHAPTER 5 --- CHARACTERIZATION OF ARDRA UNCLASSIFIABLE ACINETOBACTERS (AUA) BY FLUORESCENT AMPLIFIED FRAGMENT LENGTH POLYMORPHISM (FAFLP) METHOD --- p.98 / Chapter 5.1 --- Results / Chapter 5.1.1 --- Assessment of robustness and reproducibility --- p.98 / Chapter 5.1.2 --- Construction of the database with the reference strains --- p.104 / Chapter 5.1.2 --- Characterization of the representative AUA groups --- p.108 / Chapter 5.2 --- Discussion / Chapter 5.2.1 --- "Evaluation of robustness, reproducibility and discriminatory power" --- p.116 / Chapter 5.2.2 --- Possible genospeciation of the representative AUA groups --- p.120 / Chapter 5.2.3 --- Merits and limitations --- p.122 / Chapter CHAPTER 6 --- CHARACTERIZATION OF ARDRA UNCLASSIFABLE ACINETOBACTERS (AUA) BY PHENOTYPIC METHODS --- p.125 / Chapter 6.1 --- Results Characterization of the representative AUA groups --- p.125 / Chapter 6.2 --- Discussion / Chapter 6.2.1 --- Possible genospeciation of the representative AUA groups --- p.134 / Chapter 6.2.2 --- Limitations in classification of Acinetobacter species at genomic species level --- p.135 / Chapter CHAPTER 7 --- RELATEDNESS OF ISOLATES WITHIN THE SAME AUA GROUPS --- p.139 / Chapter 7.1 --- Results Typing results of the studied AUA groups by ERIC method --- p.139 / Chapter 7.2 --- Discussion Relatedness of the isolates within the same AUA group --- p.146 / Chapter CHAPTER 8 --- GENERAL DISCUSSION --- p.148 / Chapter 8.1 --- Possible genospeciation of the representative AUA groups --- p.150 / Chapter 8.2 --- "Comparison of ARDRA, tDNA fingerprinting, FAFLP and phenotypic methods" --- p.154 / Chapter 8.3 --- Conclusion and significance of the AUA groups studied --- p.158 / Chapter 8.4 --- Future work --- p.159 / REFERENCES --- p.160 / APPENDIX --- p.176
Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_323648 |
Date | January 2001 |
Contributors | Chu, Ka-yi., Chinese University of Hong Kong Graduate School. Division of Microbiology. |
Source Sets | The Chinese University of Hong Kong |
Language | English, Chinese |
Detected Language | English |
Type | Text, bibliography |
Format | print, xvi, 183 leaves : ill. ; 30 cm. |
Coverage | Hong Kong |
Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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