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Gewählte Doctoral Thesis:

Souzan Sanoussi (2012): DEVELOPMENT OF MICROARRAYS FOR THE MOLECULAR TYPING OF SALMONELLA ENTERICA SSP. ENTERICA.
Doctoral Thesis - Institut für Bodenforschung (IBF), BOKU-Universität für Bodenkultur, pp 131. UB BOKU obvsg

Abstract:

Identification of Salmonella enterica serovars is traditionally accomplished via immunological methods. For subtyping within serovars, phagetyping is complemented by molecular methods. Primary strain isolation and traditional serotyping as well as further conventional, phenotypic phagetyping are time-consuming and require considerable routine and experience in classification. The availability of high-quality antisera sets for serotyping and the envisaged replenishment of phage stocks are a significant burden and bottleneck. A microarray for serotyping, based on two housekeeping and two virulence marker genes (atpD, gyrB, fliC and fljB), has been previously developed by Austrian Institute of Technology (AIT) for the detection and identification of most prevalent Salmonella serovars in Austria and the UK. For further classification beyond serotyping, a novel microarray-based subtyping method based on the Diversity Arrays Technology (DArT) has recently been introduced. The DArT platform screens a large number of gDNA fragments to identify sequence-ready diagnostic markers. The first part of the present thesis is centered on the further improvement of the microarray based serotyping method. Major changes of the original technical procedure were the application of a multiplex PCR for the four targeted genes and the introduction of an alternative, more sensitive labeling method (SSELO, sequence specific end labeling of oligonucleotides). This resulted in lower consumables costs and reductions of hands-on time and total processing time. In addition, the geographical coverage of the array was extended to Greece and Denmark. As the second part of the thesis, the DArT subtyping method was applied to the analysis of a broad collection of S. Enteritidis strains. The thesis illustrates the technology underlying both microarray applications, and explains how the diagnostic tools may be useful in epidemiological investigations for achieving Salmonella strain identification.

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