Molecular genetic testing for celiac disease-associated HLA alleles

Celiac disease is an autoimmune disease, which is characterised by intolerance of gliadin creating a part of gluten. Its intolerance leads to a chronic inflammatory response in the mucous membrane of small intestine, followed by malabsorption characterized by chronic diarrhoea, greasy stool and failure to thrive. Celiac disease is a multifactorial disease, caused by external environment factors and genetic factors. Genetic predisposition is given by HLA alleles HLA-DQ2 (DQA1*05/DQB1*02) or DQ8 (DQA1*0301/DQB1*0302) or HLA-DRB1*04. This disease has got a set of unspecified clinical symptoms, that is why most of the patients are not diagnosed at all. Estimated count of the ill people is up to 50 000, however real patients are only 15 % of them. The aim of the theoretical part of the bachelor thesis was to sum up the findings about the celiac disease and molecular genetic methods based on PCR. In the theoretical part about the celiac disease, I deal mainly with the description, types, diagnostics, pathogenesis and also with the diseases associated with celiac disease. Moreover, I dealt with the description and the nomenclature of HLA system and its association with the diseases. In the end of the theoretical part, I dealt with molecular genetic methods based on PCR (PCR-SSP and RT-PCR). I carried out the practical part of the thesis in a genetic laboratory at the Faculty of Health and Social Sciences, at the Faculty of Agriculture and at GENLABS, s.r.o. in Budweis. I dealt with the HLA typisation of alleles associated with the celiac disease. I did the examination by means of two methods - PCR-SSP method with the help of HISTO TYPE Celiac Disease kit and RT-PCR method with the help of EliGene? Coeliac RT kit (DQ2, DQ8, DRB4). The aim of the work was to embrace correct laboratory practice in a genetic laboratory. Then also set and optimize a commercially sold kit for an examination of HLA alleles associated with the celiac disease on the PCR-SSP principle. Examine the already examined patients by means of RT-PCR method. Compare both methods on the terms of accordance and difficulty