Multiplex HLA-DR-DQ genotyping : for genetic epidemiology and clinical risk assessment

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Multiplex HLA-DR-DQ genotyping : for genetic epidemiology and clinical risk assessment

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dc.contributor.author Lavant, Ewa
dc.date.accessioned 2012-03-20T13:24:21Z
dc.date.available 2012-03-20T13:24:21Z
dc.date.issued 2012 en_US
dc.identifier.citation 53 s.
dc.identifier.isbn 978-91-7104-431-0 en_US
dc.identifier.issn 1653-5383 en_US
dc.identifier.uri http://hdl.handle.net/2043/13570
dc.description.abstract The human leukocyte antigens (HLA) are highly polymorphic cell surface proteins encoded in the major histocompatibility complex (MHC) region on chromosome 6. The HLA system has been well known as transplantation antigens but the primary biological role of the HLA molecules is regulation of immune response by presenting peptide fragments to T-lymphocytes. As regulators of immune responses the HLA molecules are also of importance for susceptibility to several autoimmune and inflammatory diseases. Genotyping of these loci is therefore significant in research targeting the mechanisms of HLA associated diseases, in exploring new epidemiological associations between HLA and specific disease, and as a clinical tool for risk assessment for diseases with well defined associations. Although several commercial HLA genotyping methods are available, many require multiple steps, have low throughput and high cost. The aim of the work within this dissertation was to develop a robust, costeffective method for HLA-DRB1, -DQA1 and -DQB1 genotyping suitable for use in an epidemiological context and clinical investigation. The method was optimized with specific focus on risk alleles for type 1 diabetes mellitus and celiac disease, two autoimmune disorders with significant impact on public health. By combining PCR with sequence specific primers (PCR-SSP), product separation by capillary gel electrophoresis and fluorescence detection in the developed method, all three loci could be analyzed in a single step, resulting in low reagent cost and fast turnaround time. This in combination with the low total consumption of DNA template allows the method to be used in epidemiological studies. 10 A simplified version of the developed method is currently used for clinical risk assessment for celiac disease when histological and/or serologic results are ambiguous in investigated subjects or when a gluten-free diet has been initiated before diagnostic tests have been performed. The low cost of this newly developed method has enabled HLA typing as a tool in screening programs for high-risk groups, such as individuals with Down syndrome or type 1 diabetes, to preclude the risk for celiac disease and thus avoid periodic screening for auto-antibodies. This method is also used to analyze samples from children all over Sweden with newly diagnosed diabetes in the Better Diabetes Diagnosis project. The developed method was also used in two explorative association studies not related to type 1 diabetes or celiac disease. In one study the association between HLA-DRB1, -DQA1 and -DQB1 and acute myocardial infarction was investigated showing only weak associations. In the second study the HLA-DR-DQ haplotype effect on developing chronic pain after inguinal hernia surgery was explored demonstrating an HLA dependent risk of developing pain
dc.language.iso eng en_US
dc.publisher Malmö University. Faculty of Health and Society en_US
dc.relation.ispartofseries Doctoral Dissertation;2012:3
dc.relation.haspart Lavant EH, Carlson JA. A new automated HLA-typing strategy to identify DR-DQ risk alleles for celiac disease and Type 1 Diabetes Mellitus. Clin Chem Lab Med. 2009;47:1489-95.
dc.relation.haspart Lavant EH, Agardh D, Nilsson A, Carlson JA. A new PCR-SSP method for HLA DR-DQ risk assessment for celiac disease. Clin Chim Acta. 2011;412:782-4.
dc.relation.haspart Björkbacka H, Lavant EH, Nordin Fredrikson G, Melander O, Berglund G, Carlson J, Nilsson J. Weak associations between human leukocyte antigen (HLA) genotype and acute myocardial infarction. J Intern Med. 2010;268:50-8.
dc.relation.haspart Dominguez C, Kalliomäki M, Gunnarsson U, Sandblom G, Kockum I, Lavant E, Olsson T, Nyberg F, Gordh T, Piehl F. The DRB1*04-DQB1*03:02 HLA haplotype is associated with increased risk of chronic pain after inguinal hernia surgery. Manuscript submitted to Pain.
dc.subject HLA-DRB1
dc.subject HLA-DQB1
dc.subject HLA-DQA1
dc.subject PCR-SSP
dc.subject type 1 diabetes
dc.subject celiac disease
dc.subject autoimmune disease
dc.subject capillary gel electrophoresis
dc.subject genotyping
dc.subject.classification Medicine en_US
dc.title Multiplex HLA-DR-DQ genotyping : for genetic epidemiology and clinical risk assessment en_US
dc.type Doctoral Thesis
dc.identifier.paperprint 1 en_US
dc.contributor.department Malmö University. Faculty of Health and Society en
dc.description.other en_US
dc.subject.srsc Research Subject Categories::MEDICINE en_US
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