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

Clinical Chemistry and Laboratory Medicine : CCLM
Ewa H Lavant, Joyce A Carlson

Abstract

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. 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. 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. 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.

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Citations

Jun 22, 2011·Acta Diabetologica·Antonio PicarelliPatrizia Gargiulo
Jun 18, 2015·Scandinavian Journal of Clinical and Laboratory Investigation·Flemming LundGyörgy Sölétormos
Jan 12, 2011·Clinica Chimica Acta; International Journal of Clinical Chemistry·Ewa H LavantJoyce A Carlson
Nov 16, 2010·Journal of Diabetes·Li Zhang, George S Eisenbarth
Sep 23, 2014·Critical Reviews in Clinical Laboratory Sciences·Vilte E BarakauskasMathew P Estey
May 4, 2016·Journal of Diabetes and Its Complications·Amit AkirovIlan Shimon
May 7, 2013·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Linda AkermanRosaura Casas
Apr 17, 2012·Clinical Chemistry and Laboratory Medicine : CCLM·Karsten ConradMartin W Laass
Oct 10, 2014·BMC Gastroenterology·Hanna BragdeJan Söderman
Jan 16, 2019·Current Rheumatology Reviews·Elham RajaeiSeyed Mohammad Sadegh Pezeshki
Oct 13, 2020·Scandinavian Journal of Clinical and Laboratory Investigation·Flemming LundSøren Kristiansen

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