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A new automated human leukocyte antigen genotyping strategy to identify DR-DQ risk alleles for celiac disease and type 1 diabetes mellitus
-
Ewa H. Lavant
Veröffentlicht/Copyright:
1. Dezember 2009
Abstract
Background: The risk for type 1 diabetes mellitus (T1DM) and celiac disease (CD) is related to human leukocyte antigen (HLA) DQA1, DQB1 and DRB1 loci. Unfortunately, HLA typing has been too difficult and costly for frequent use. Automated genotyping focused on risk alleles could provide access to HLA typing in diagnostic evaluations, epidemiological screening and contribute to preventive strategies.
Methods: A sequence specific primer amplification method requiring a total of four PCR reactions, one restriction enzyme digestion and a single electrophoretic step provides low to medium resolution typing of DQA1, DQB1 and DRB1 using Applied Biosystems 3730 DNA analyzer. The method was validated using 261 samples with genotypes determined using a reference method.
Results: Specific fluorescent DQA1, DQB1 and DRB1 amplicons were of expected size. Concordance with the reference method was 100% for DQA1 and DQB1 alleles and 99.8% for DRB1 alleles.
Conclusions: We have developed a high throughput HLA typing method that accurately distinguishes risk alleles for T1DM and CD. This method allows screening of several thousand samples per week, consuming 32 ng of DNA template, low reagent volumes and minimal time for data review.
Clin Chem Lab Med 2009;47:1489–95.
Keywords: automation; celiac disease; HLA-DR-DQ-genotyping; PCR-SSP (sequence specific priming); type 1 diabetes mellitus
Received: 2009-6-18
Accepted: 2009-9-1
Published Online: 2009-12-01
Published in Print: 2009-12-01
©2009 by Walter de Gruyter Berlin New York
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Schlagwörter für diesen Artikel
automation;
celiac disease;
HLA-DR-DQ-genotyping;
PCR-SSP (sequence specific priming);
type 1 diabetes mellitus
Artikel in diesem Heft
- Review
- Multiple gene interaction and modulation of hemostatic balance
- Minireview
- Appropriate utilization of clinical laboratory tests
- Genetics and Molecular Diagnostics
- Quantitation of RNA decay in dried blood spots during 20 years of storage
- ROS1 Asp2213Asn polymorphism is not associated with coronary artery disease in a Greek case-control study
- Association of glutathione-S-transferase polymorphisms with atopic dermatitis risk in preschool age children
- Haplotype-based association of regulator of G-protein signaling 5 gene polymorphisms with essential hypertension and metabolic parameters in Chinese
- A new automated human leukocyte antigen genotyping strategy to identify DR-DQ risk alleles for celiac disease and type 1 diabetes mellitus
- General Clinical Chemistry and Laboratory Medicine
- Serum thyrotropin and free thyroxine reference ranges as defined in a disease-free sample of French middle-aged adults
- Screening for M-proteinemia: serum protein electrophoresis and free light chains compared
- Temporal profile and clinical significance of serum neuron-specific enolase and S100 in ischemic and hemorrhagic stroke
- Salivary neuron specific enolase: an indicator for neuronal damage in patients with ischemic stroke and stroke-prone patients
- Antibodies to mutated citrullinated vimentin and antibodies to cyclic citrullinated peptides in juvenile idiopathic arthritis
- Age-associated developmental changes in the activated partial thromboplastin time (APTT) and causes of prolonged APTT values in healthy Chinese children
- Nucleated red blood cells and soluble transferrin receptor in thalassemia syndromes: relationship with global and ineffective erythropoiesis
- Serum chitotriosidase enzyme activity in patients with Crimean-Congo hemorrhagic fever
- Preanalytical mistakes in samples from primary care patients
- Should kidney tubular markers be adjusted for urine creatinine? The example of urinary cystatin C
- Validation and Outcome Studies
- Development and analytical performance evaluation of an automated chemiluminescent immunoassay for pro-gastrin releasing peptide (ProGRP)
- Development and validation of a liquid chromatography-tandem mass spectrometry assay for serum 25-hydroxyvitamin D2/D3 using a turbulent flow online extraction technology
- Letters to the Editor
- Lack of association between eNOS Glu298Asp gene polymorphism and carotid atherosclerosis in a Serbian population
- Falsely elevated troponin I attributed to collection tubes using the Vitros ECiQ system
- Processing effects and storage conditions on A Disintegrin And Metalloprotease (ADAM12s), a maternal serum marker for adverse pregnancy outcome
- Serum γ-glutamyltransferase: linking together environmental pollution, redox equilibria and progression of atherosclerosis?
- The impact factor for evaluating scientists: the good, the bad and the ugly
- Acknowledgement
- Acknowledgement
- Contents
- Contents, Volume 47, 2009
- Author Index
- Author Index
- Subject Index
- Subject Index
