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Medium-high resolution electrochemical genotyping of HLA-DQ2/DQ8 for detection of predisposition to coeliac disease

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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.

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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Acknowledgement

This work was carried out with the financial support from the Commission of the European Communities, specific RDT program Coeliac Disease Management Monitoring and Diagnosis using Biosensors and an Integrated Chip System, CD-MEDICs, FP7-2007-ICT-1-216031. V.B. acknowledges the European Union’s Seventh Framework Program (FP7/2007-2013) under grant agreement no. PIGF-GA-2008-220928 for financial support. H.J. thanks Universitst Rovira i Virgili for the doctoral scholarship. The authors thank Dr. Markku Heikkinen, MD, PhD (Kuopio University Hospital, Kuopio, Finland) for blood samples of Finnish celiac disease patients and Dr. Taina Jaatinen, PhD, Dr. Katri Haimila, PhD, and Juha Peräsaari, MS (Finnish Red Cross Blood Service, Helsinki, Finland) for HLA tissue typing expertise.

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Author notes

  1. Valerio Beni

    Present address: Biosensors & Bioelectronics Centre, Department of Physics, Chemistry and Biology (IFM), Linköping University, 58183, Linköping, Sweden

Authors and Affiliations

  1. Departament d’Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans, 26, 43007, Tarragona, Spain

    Hamdi Joda, Valerio Beni, Ioanis Katakis & Ciara K. O’Sullivan

  2. Finnish Red Cross Blood Service, Kivihaantie 7, 00310, Helsinki, Finland

    Noora Alakulppi & Jukka Partanen

  3. TATAA Biocenter AB, Odinsgatan 28, 41103, Göteborg, Sweden

    Kristina Lind & Linda Strömbom

  4. Institut für Mikrotechnik Mainz GmbH, Carl-Zeiss-Strasse 18-20, 55129, Mainz, Germany

    Daniel Latta & Julian Höth

  5. Institucio Catalana de Recerca i Estudis Avançats, Passeig Lluis Companys 23, 08010, Barcelona, Spain

    Ciara K. O’Sullivan

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  1. Hamdi Joda
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Correspondence to Valerio Beni or Ciara K. O’Sullivan.

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Joda, H., Beni, V., Alakulppi, N. et al. Medium-high resolution electrochemical genotyping of HLA-DQ2/DQ8 for detection of predisposition to coeliac disease. Anal Bioanal Chem 406, 2757–2769 (2014). https://doi.org/10.1007/s00216-014-7650-x

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  • DOI: https://doi.org/10.1007/s00216-014-7650-x

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