Medium-high resolution electrochemical genotyping of HLA-DQ2/DQ8 for detection of predisposition to coeliac disease |
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Authors: | Hamdi Joda Valerio Beni Noora Alakulppi Jukka Partanen Kristina Lind Linda Strömbom Daniel Latta Julian Höth Ioanis Katakis Ciara K O’Sullivan |
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Institution: | 1. Departament d’Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Pa?sos Catalans, 26, 43007, Tarragona, Spain 6. Biosensors & Bioelectronics Centre, Department of Physics, Chemistry and Biology (IFM), Link?ping University, 58183, Link?ping, Sweden 2. Finnish Red Cross Blood Service, Kivihaantie 7, 00310, Helsinki, Finland 3. TATAA Biocenter AB, Odinsgatan 28, 41103, G?teborg, Sweden 4. Institut für Mikrotechnik Mainz GmbH, Carl-Zeiss-Strasse 18-20, 55129, Mainz, Germany 5. Institucio Catalana de Recerca i Estudis Avan?ats, Passeig Lluis Companys 23, 08010, Barcelona, Spain
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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. Figure Electrode array and schematic of the proposed detection approach for the medium to high resolution electrochemical genotyping of alleles associated to Coeliac disease |
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