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

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, DOB1*02) and/or HLA-DQ8 (DQA1*03, DOB1*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 CID risk evaluation and of a second kit, namely DQ-CD Zygosis, for DOB1*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 CID. Molecular HLA testing is considered a valid support in the confirmation/exclusion of CID, especially in high-risk groups, such as CID relatives, or when serological and histological data are ambiguous.