Abstract
The risk for celiac disease (CD) is clearly related to specific HLA DQA1 and DQB1 alleles, but HLA typing is often considered too costly for frequent use.
Here we present a method using sequence-specific primed PCR (PCR-SSP) for HLA-DR-DQ genotyping optimized for capillary electrophoresis on Applied Biosystems 3130xl Genetic Analyzer. Requiring a total of three PCR reactions and a single electrophoretic step, this method reduces the reagent expenses and technical time for directed HLA typing to distinguish risk alleles for CD, with a sufficient throughput for large-scale screening projects.
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References
Farrell RJ, Kelly CP (2002) Celiac sprue. N Engl J Med 346:180–188
Green PH, Cellier C (2007) Celiac disease. N Engl J Med 357:1731–1743
Dube C et al (2005) The prevalence of celiac disease in average-risk and at-risk Western European populations: a systematic review. Gastroenterology 128:S57–S67
Fasano A et al (2003) Prevalence of celiac disease in at-risk and not-at-risk groups in the United States: a large multicenter study. Arch Intern Med 163:286–292
Megiorni F et al (2009) HLA-DQ and risk gradient for celiac disease. Hum Immunol 70:55–59
Sollid LM, Lie BA (2005) Celiac disease genetics: current concepts and practical applications. Clin Gastroenterol Hepatol 3:843–851
Louka AS, Sollid LM (2003) HLA in coeliac disease: unravelling the complex genetics of a complex disorder. Tissue Antigens 61:105–117
Margaritte-Jeannin P et al (2004) HLA-DQ relative risks for coeliac disease in European populations: a study of the European Genetics Cluster on Coeliac Disease. Tissue Antigens 63:562–567
Vader W et al (2003) The HLA-DQ2 gene dose effect in celiac disease is directly related to the magnitude and breadth of gluten-specific T cell responses. Proc Natl Acad Sci USA 100:12390–12395
Karell K et al (2003) HLA types in celiac disease patients not carrying the DQA1*05-DQB1*02 (DQ2) heterodimer: results from the European Genetics Cluster on Celiac Disease. Hum Immunol 64:469–477
Kaukinen K et al (2002) HLA-DQ typing in the diagnosis of celiac disease. Am J Gastroenterol 97:695–699
Pietzak MM et al (2009) Stratifying risk for celiac disease in a large at-risk US population using HLA alleles. Clin Gastroenterol Hepatol 7:966–971
Karinen H et al (2006) HLA genotyping is useful in the evaluation of the risk for coeliac disease in the 1st-degree relatives of patients with coeliac disease. Scand J Gastroenterol 41:1299–1304
Anthropology/Allele Frequencies database at dbMHC at the NCBI homepage. Available at: http://www.ncbi.nlm.nih.gov/gv/mhc/main.fcgi?cmd=init
Lavant EH et al (2011) A new PCR-SSP method for HLA DR-DQ risk assessment for celiac disease. Clin Chim Acta 412:782–784
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Parts of the text have been modified from Clinica Chimica Acta with permission from Elsevier publishers (15).
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Lavant, E.H., Carlson, J. (2013). HLA DR-DQ Genotyping by Capillary Electrophoresis for Risk Assessment for Celiac Disease. In: Phillips, T., Kalish, H. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 919. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-029-8_26
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DOI: https://doi.org/10.1007/978-1-62703-029-8_26
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-028-1
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