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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

SCImago Journal Rank (SJR) 2016: 1.000
Source Normalized Impact per Paper (SNIP) 2016: 1.112

Online
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1437-4331
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Volume 47, Issue 12 (Dec 2009)

Issues

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
  • Department of Biomedical Laboratory Science, Faculty of Health and Society, Malmö University, Malmö, Sweden
  • Labmedicine Skåne, Clinical Chemistry in Malmö, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Joyce A. Carlson
  • Labmedicine Skåne, Clinical Chemistry in Malmö, Sweden
  • Labmedicine Skåne, Clinical Chemistry in Lund, Sweden
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar

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

About the article

Corresponding author: Ewa Lavant, Labmedicine Skåne, Clinical Chemistry in Malmö, Entrance 71, University Hospital in Malmö, 205 02 Malmö, Sweden Phone: +46 40 33 64 47/+46 762 67 88 91, Fax: +46 40 33 62 86,


Received: 2009-06-18

Accepted: 2009-09-01

Published in Print: 2009-12-01


Citation Information: Clinical Chemistry and Laboratory Medicine, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2009.346.

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©2009 by Walter de Gruyter Berlin New York. Copyright Clearance Center

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