A new automated human leukocyte antigen genotyping strategy to identify DR-DQ risk alleles for celiac disease and type 1 diabetes mellitus

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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

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Publication Article, peer reviewed scientific
Title A new automated human leukocyte antigen genotyping strategy to identify DR-DQ risk alleles for celiac disease and type 1 diabetes mellitus
Author(s) Lavant, Ewa ; Carlson, Joyce
Date 2009
English 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.
DOI http://dx.doi.org/10.1515/CCLM.2009.346 (link to publisher's fulltext)
Publisher Walter De Gruyter
Host/Issue Clinical Chemistry and Laboratory Medicine;12
Volume 47
ISSN 1434-6621
Pages 1489-9
Language eng (iso)
Subject(s) HLA-DR-DQ-genotyping
celiac disease
type 1 diabetes mellitus
Medicine
Research Subject Categories::MEDICINE
Handle http://hdl.handle.net/2043/9041 (link to this page)

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