|
|
Molecular detection of selected gene polymorphisms related to nutrition (nutrichip validation)
TURKOVÁ, Kateřina
Lactose intolerance is the most common food intolerance in the world. Individuals with lactose intolerance are unable to produce the enzyme lactase in the small intestine, which makes it possible to break down the lactose contained in dairy products. Insufficient lactase production may be genetically determined. Two single nucleotide polymorphisms responsible for the persistence of lactase activity in adulthood have been found in the European population. Celiac disease is one of the autoimmune diseases that mainly affects the mucous membrane of the small intestine. The disease is characterized by intolerance to gliadin, which is part of gluten. Intolerance leads to chronic inflammation of the small intestinal mucosa, leading to chronic diarrhea, fatty stools, vomiting and fatigue. The development of celiac disease is conditioned by the presence of a genetic predisposition. Genetic predisposition is linked to HLA system alleles. Specifically, these are the HLA-DQ2 and HLA-DQ8 haplotypes.
|
|
|
Quality control of molecular biology methods in transplantation medicine
Kotrbatý, Jiří ; Kolesár, Libor (advisor) ; Schierová, Michaela (referee)
This work is focused on the description of quality control of molecular-biological methods used in transplant medicine, ie methods of HLA typing. Quality control is part of the quality management system. It consists of several components: validation, verification, internal quality control and external quality assessment. Each medical laboratory, including laboratories engaged in HLA typing, these elements must be applied to its routine operation. Validation is done before the introduction of methods and verifies whether the method is correct for the intended use. Verification is to verify that the validated method is used correctly in laboratory conditions. Internal quality control is based on an analysis of known positive and negative samples and evaluate the accuracy and precision of the whole analytical process. The external evaluation of the quality of the different laboratories analyze the same sample and the results are compared to each other. Key words: HLA typing, validation, verification, internal quality control, external quality assessment
|
|
|
Study of HLA antigens and KIR genes in a donors and recipients of bone marrow
Kroulíková, Zuzana ; Vraná, Milena (advisor) ; Froňková, Eva (referee)
HLA and KIR genes are highly polymorphic regions within the human genome. Protein products of these genes play a critical role in hematopoietic stem cell transplantation. Genetic HLA match is a major barrier to engraftment and influences the outcome of this therapy. Therefore it is necessary to genotype donors and recipients selected for hematopoietic stem cell transplantation. Today HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 genes are tested by modifications of polymerase chain reaction or by sequence-based typing methods on the level of high- or low-resolution. Donors registered in bone marrow registries are selected on the basis of a 10/10 match. Donors KIR genotype leads to a better outcome, to relapse-free survival and overall survival in treatment of patients with acute myeloid leukemia. A better protection against relapse is achieved by Cen-B/B donor haplotype. Therefore KIR typing by polymerase chain reaction is used and the genotype is compared with the IMGT/KIR database by an online B- content calculator. Donors are divided in groups according to their genotype and their influence on the success of treatment for acute myeloid leukemia. The study of polymorphic systems and the development of genotyping donors and recipients significantly improve the outcome of hematopoietic stem cell...
|
|
|
Examination of genetic predispositions for celiac disease in a family with a history of diabetes mellitus 1. Type
KUČEROVÁ, Klára
Celiac disease is one of the autoimmune disorders that mainly affects the mucous membrane of the small intestine. The disease is characterized by intolerance to gliadin, which forms part of gluten. The intolerance leads to chronic inflammation of the mucous membrane of the small intestine, causing chronic diarrhea, adipose feces, vomiting, fatigue, and last but not least also weight loss. Type I. diabetes mellitus is an autoimmune metabolic disease demonstrated by hyperglycemia (increased level of blood glucose) which is a result of the insufficient effect of insulin. Type I. DM can occur at individuals as an independent disease or combined with other autoimmune diseases such as celiac disease. While genetic predisposition is a condition for the development of celiac disease, the development is influenced to a greater extent by external factors, for example environmental elements. Genetic predisposition is bound to alleles of the HLA system. This concerns in particular the HLA-DQ2 and HLA-DQ8 haplotypes. Due to various clinical symptoms, it is very difficult to diagnose the disease in some cases, and many patients are not diagnosed with the disease at all. The aim of the theoretical part of the bachelor thesis is to study and then summarize findings concerning celiac disease and type I. diabetes mellitus (DM I). To a great extent, the thesis addresses the topics of diabetes, celiac disease, and the HLA system. As for celiac disease, the thesis focuses on its characteristics, diagnostics, treatment, and inheritance. In the case of diabetes, the thesis deals with its individual types, treatment and also the genetic predispositions. The conclusion of the thesis describes the HLA system itself and shows the importance of the association between the type I. diabetes mellitus and celiac disease. The practical part of the thesis was conducted in the GENLABS genetics laboratory in České Budějovice. In this laboratory, genetic predispositions to celiac disease were examined by using the real-time PCR method. The examination is based on the HLA typing of the risk alleles by means of the commercially produced EliGene? Coeliac RT kit (DQ2, DQ8, DR4). The goal of the practical part is to master the basic laboratory methods such as the DNA isolation from the peripheral blood and buccal swab, to measure the DNA concentration, to adopt the real-time PCR method that serves to determine the HLA risk haplotype and to analyze gained results.
|
|
|
Molecular genetic testing for celiac disease-associated HLA alleles
SALZMANOVÁ, Kateřina
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.
|
|
|
Study of HLA antigens and KIR genes in a donors and recipients of bone marrow
Kroulíková, Zuzana ; Vraná, Milena (advisor) ; Froňková, Eva (referee)
HLA and KIR genes are highly polymorphic regions within the human genome. Protein products of these genes play a critical role in hematopoietic stem cell transplantation. Genetic HLA match is a major barrier to engraftment and influences the outcome of this therapy. Therefore it is necessary to genotype donors and recipients selected for hematopoietic stem cell transplantation. Today HLA-A, HLA-B, HLA-C, HLA-DRB1 and HLA-DQB1 genes are tested by modifications of polymerase chain reaction or by sequence-based typing methods on the level of high- or low-resolution. Donors registered in bone marrow registries are selected on the basis of a 10/10 match. Donors KIR genotype leads to a better outcome, to relapse-free survival and overall survival in treatment of patients with acute myeloid leukemia. A better protection against relapse is achieved by Cen-B/B donor haplotype. Therefore KIR typing by polymerase chain reaction is used and the genotype is compared with the IMGT/KIR database by an online B- content calculator. Donors are divided in groups according to their genotype and their influence on the success of treatment for acute myeloid leukemia. The study of polymorphic systems and the development of genotyping donors and recipients significantly improve the outcome of hematopoietic stem cell...
|
|
|
Quality control of molecular biology methods in transplantation medicine
Kotrbatý, Jiří ; Kolesár, Libor (advisor) ; Schierová, Michaela (referee)
This work is focused on the description of quality control of molecular-biological methods used in transplant medicine, ie methods of HLA typing. Quality control is part of the quality management system. It consists of several components: validation, verification, internal quality control and external quality assessment. Each medical laboratory, including laboratories engaged in HLA typing, these elements must be applied to its routine operation. Validation is done before the introduction of methods and verifies whether the method is correct for the intended use. Verification is to verify that the validated method is used correctly in laboratory conditions. Internal quality control is based on an analysis of known positive and negative samples and evaluate the accuracy and precision of the whole analytical process. The external evaluation of the quality of the different laboratories analyze the same sample and the results are compared to each other. Key words: HLA typing, validation, verification, internal quality control, external quality assessment
|
|
|
Molecular biology methods and their use for HLA antigen polymorphism determination
Brožová, Jitka ; Slavčev, Antonij (advisor) ; Trošan, Peter (referee)
HLA (Human Leukocyte Antigen) is a gene segment localized on the short arm of chromosome 6. This segment consists of three regions, I. and II. region encode HLA class I and HLA class II genes, in the III. region there are the complement genes, TNFα and other genes that are not evolutionary and functionally associated with HLA genes. HLA class I and class II genes encode cell-surface glycoproteins which in their binding sites present peptides to cells of the immune system. HLA genes are associated with many autoimmune and infectious diseases, although mostly the mechanisms of these associations remain unclear. HLA function as strong transplantation antigens, thus they represent a major obstacle in allogeneic organ transplantation and transplantation of hematopoietic cells. To prevent rejection of the transplanted organ, a match between the recipient and donor in the HLA antigens is necessary. Determination of donor and recipient matching is performed by HLA typing techniques. Nowadays for HLA typing in most laboratories in Europe and the USA are used methods of molecular biology. Among these methods, three main techniques based on the polymerase chain reaction (PCR) are applied, these are PCR-sequence-specific primers, PCR-sequence-specific oligonucleotide probes and PCR-sequencing-based typing....
|
guest ::
