Detection of coeliac disease predisposition using dna biosensor arrays

Identificador:  TDX:1093

Handle: https://hdl.handle.net/20.500.11797/TDX1093

Autors:  Joda, Hamdi Abdelazim Osman

Resum:
Coeliac disease (CD) is a small intestinal inflammation, affecting genetically susceptible individuals, triggered by ingestion of certain cereals. Prevalence of CD in Europe and US is about 1%. Most of CD cases remain undiagnosed for many years because of highly variable clinical spectrum and atypical presentation. The gold standard test for CD diagnosis is the small-intestinal biopsy, an invasive and expensive test. Strong relation between CD and Human Leukocyte Antigens (HLA) has been proved, as 95% of the CD patients carrying HLA DQ2 heterodimer and 5% carrying DQ8 heterodimer. DQ2 and DQ8 negative individuals have been shown to be very unlikely to develop CD. The overall objective of the thesis is to develop a rapid, ease to use, cost effective diagnostic genosensor array for DQ2/DQ8 typing as a diagnostic tool for CD predisposition. Two different methods have been investigated in parallel. In the first method, the genosensor array was developed employing the SSOP method, by designing different allele specific probes. Whilst in the second approach, the SSP technique exploiting a novel approach for the direct and rapid detection of a double stranded PCR amplicon was investigated. In both approaches, assay conditions were optimised and finally analysis of real clinical samples was performed.