title: Low-medium resolution HLA-DQ2/DQ8 typing for coeliac disease predisposition analysis by colorimetric assay.
abstract: Coeliac disease is an inflammation of the small intestine, occurring in genetically susceptible individuals triggered by the ingestion of gluten. Human Leukocyte Antigens (HLA) DQ2 and DQ8 gene have been identified as key genetic factors in coeliac disease as they are presented in almost 100 % of the patients. These genes are encoded by the combination of certain alleles in the DQA and DQB region of chromosome 6. Specifically, DQA1*05:01 and DQB1*02:01 alleles for serologically defined leukocyte antigen DQ2 cis, DQA1*05:05 and DQB1*02:02 for DQ2 trans and DQA1*03:01 and DQB1*03:02 alleles for the DQ8. Specific identification of these alleles is a challenge due to the high number of alleles that have been identified so far: 46 in the DQA region and 160 in the DQB region (as of IMGT/HLA Database 10/2011 release). In the reported work, the development of a multiplex colorimetric assay for the low to medium HLA typing of the DQ2 and DQ8 genes is presented. The optimisation of probe design and assay conditions, performed by both surface plasmon resonance and enzyme-linked oligonucleotide assay, are reported. Finally, the performances of the developed typing platform were validated by the analysis of real patient samples and HLA typing, compared with those obtained using hospital based typing technology and an excellent correlation obtained.
authors.affiliation: Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans, 26, 43007, Tarragona, Spain.
authors.forename: Hamdi
authors.initials: H
authors.lastname: Joda
authors.forename: Valerio
authors.initials: V
authors.lastname: Beni
authors.forename: Deirdre
authors.initials: D
authors.lastname: Curnane
authors.forename: Ioanis
authors.initials: I
authors.lastname: Katakis
authors.forename: Noora
authors.initials: N
authors.lastname: Alakulppi
authors.forename: Jukka
authors.initials: J
authors.lastname: Partanen
authors.forename: Kristina
authors.initials: K
authors.lastname: Lind
authors.forename: Linda
authors.initials: L
authors.lastname: Strömbom
authors.forename: Ciara K
authors.initials: CK
authors.lastname: O'Sullivan
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002446:Celiac Disease / Q000235:genetics*; D003124:Colorimetry / Q000379:methods; D015342:DNA Probes / Q000235:genetics; D020022:Genetic Predisposition to Disease; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D006683:HLA-DQ Antigens / Q000235:genetics*; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods; D012680:Sensitivity and Specificity; D020349:Surface Plasmon Resonance
chemical_list: D015342:DNA Probes; D006683:HLA-DQ Antigens; C053186:HLA-DQ2 antigen; C109756:HLA-DQ8 antigen