Application of PCR-mediated DNA typing in the molecular epidemiology of medically important microorganisms

Abstract

This thesis describes the development, application and validation of the newer DNA analysis techniques within the field of microbiological epidemiology. Emphasis is placed on the use of the polymerase chain reaction (PCR), a test-tube technique enabling the amplification of (parts of) DNA molecules to enormous amounts. By comparing the genomes of microbes, insight in the mode of dissemination of given microorganisms can be obtained. Besides, this type of laboratory procedures also allows evolutionary studies, highlighting genomic variability perse. In the first chapter the teclmology will be introduced and the function of molecular typing within a microbiological laboratory will be explained. Subsequently, present day literature describing the application of PCR in microbial epidemiology will be summarised (chapter II). Chapters III and IV provide examples of the novel PCR approaches for molecular tracking of the protozoan parasites Naeg/eria spp. and Giardia duodenalis. The fact that the application of these techniques is not restricted to free-living amoebae and intestinal parasites is demonstrated in chapters V to VIII. Monitoring spread and persistence of fungal pathogens is performed, indicating the clinical relevance of genetic screening of a diversity of fungal agents, pathogens that went through a steep rise in clinical incidence during the past ten years. The last four chapters (IX to XII) describe the technical possibilities for and the clinical implications of molecular epidemiological studies on (methicillin-resistant) Staphylococcus aureus. Finally, chapter XIII integrates data presented in the previous chapters and gives a summary of current knowhow and sketches future developments