title: Medium-high resolution electrochemical genotyping of HLA-DQ2/DQ8 for detection of predisposition to coeliac disease.
abstract: Coeliac disease is a small intestinal disorder, induced by ingestion of gluten in genetically predisposed individuals. Coeliac disease has been strongly linked to human leukocyte antigens (HLA) located on chromosome 6, with almost 100 % of coeliac disease sufferers carrying either a HLA-DQ2 or HLA-DQ8 heterodimer, with the majority carrying HLA-DQ2 encoded by the DQA1*05:01/05:05, DQB1*02:01/02:02 alleles, whereas the remaining carry the HLA-DQ8 encoded by the DQA1*03:01, DQB1*03:02 alleles. In this work, we present the development of a multiplex electrochemical genosensor array of 36 electrodes, housed within a dedicated microfluidic platform and using a total of 10 sequence-specific probes for rapid medium-high resolution HLA-DQ2/DQ8 genotyping. An evaluation of the selectivity of the designed probes was carried out with the target sequences and 44 potentially interfering alleles, including single base mismatch differentiations; good selectivity was demonstrated. The performance of the electrochemical genosensor array was validated, analyzing real human samples for the presence of HLA-DQ2/DQ8 alleles, and compared with those obtained using laboratory-based HLA typing, and an excellent correlation was 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: 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: Daniel
authors.initials: D
authors.lastname: Latta
authors.forename: Julian
authors.initials: J
authors.lastname: Höth
authors.forename: Ioanis
authors.initials: I
authors.lastname: Katakis
authors.forename: Ciara K
authors.initials: CK
authors.lastname: O'Sullivan
mesh_terms: D000483:Alleles; D002446:Celiac Disease / Q000175:diagnosis* / Q000235:genetics*; D004198:Disease Susceptibility; D055664:Electrochemical Techniques / Q000379:methods*; D005838:Genotype; D006683:HLA-DQ Antigens / Q000032:analysis / Q000235:genetics*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*
chemical_list: D006683:HLA-DQ Antigens; C053186:HLA-DQ2 antigen; C109756:HLA-DQ8 antigen