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Clinical Chemistry and Laboratory Medicine (CCLM)

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

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

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A rapid and sensitive method to detect specific human lymphocyte antigen (HLA) class II alleles associated with celiac disease

Francesca Megiorni1 / Barbara Mora2 / Margherita Bonamico3 / Raffaella Nenna4 / Mariarosaria Di Pierro5 / Carlo Catassi6 / Sandro Drago7 / Maria Cristina Mazzilli8

1Department of Experimental Medicine, “Sapienza” University of Rome, Rome, Italy

2Department of Experimental Medicine, “Sapienza” University of Rome, Rome, Italy

3Department of Pediatrics, “Sapienza” University of Rome, Rome, Italy

4Department of Pediatrics, “Sapienza” University of Rome, Rome, Italy

5Bionat Italia S.r.l., Palermo, Italy

6Department of Pediatrics, University of Ancona, Ancona, Italy

7Bionat Italia S.r.l., Palermo, Italy

8Department of Experimental Medicine, “Sapienza” University of Rome, Rome, Italy

Corresponding author: Maria Cristina Mazzilli, Department of Experimental Medicine, “Sapienza” University of Rome Viale Regina Elena, 324, 00161 Rome, Italy Phone: +39-06-4451286, Fax: +39-06-4454820

Citation Information: Clinical Chemical Laboratory Medicine. Volume 46, Issue 2, Pages 193–196, ISSN (Online) 14374331, ISSN (Print) 14346621, DOI: 10.1515/CCLM.2008.049, December 2007

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Background: Celiac disease (CD) is a complex disorder triggered by gluten affecting genetically predisposed individuals. More than 90% of patients carry human lymphocyte antigen (HLA)-DQ2 (DQA1*05, DQB1*02) and/or HLA-DQ8 (DQA1*03, DQB1*0302). We propose the use of the DQ-CD Typing kit that allows identification of the HLA class II alleles (DQA1*0201,*03,*05, DQB1*02,*0302, DRB1*03,*04,*07) selected to be informative in the CD risk evaluation and of a second kit, namely DQ-CD Zygosis, for DQB1*02 homozygosity determination.

Methods: The study was performed on a cohort of 100 individuals previously HLA typed with commercial kits. Fresh blood or previously extracted DNA was amplified in a unique PCR program using allele-specific primers and visualized on agarose gel.

Results: DNA amplification yielded strong and clear products without non specific signals or ghost bands. All the samples showed the expected alleles in accordance with the previous HLA typing.

Conclusions: The DQ-CD Typing and Zygosis kits are fast, simple, economical and accurate tools that can be used to determinate the HLA-DQ2/DQ8 status in laboratory practice addressed for the diagnosis of CD. Molecular HLA testing is considered a valid support in the confirmation/exclusion of CD, especially in high-risk groups, such as CD relatives, or when serological and histological data are ambiguous.

Clin Chem Lab Med 2008;46:193–6.

Keywords: celiac disease; DQ2/DQ8 molecules; human lymphocyte antigen (HLA) typing; sequence specific primer-polymerase chain reaction (SSP-PCR)

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