Joda Hamdi, Beni Valerio, Alakulppi Noora, Partanen Jukka, Lind Kristina, Strömbom Linda, Latta Daniel, Höth Julian, Katakis Ioanis, O'Sullivan Ciara K
Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans, 26, 43007, Tarragona, Spain.
Anal Bioanal Chem. 2014 May;406(12):2757-69. doi: 10.1007/s00216-014-7650-x. Epub 2014 Mar 15.
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 DQA105:01/05:05, DQB102:01/02:02 alleles, whereas the remaining carry the HLA-DQ8 encoded by the DQA103:01, DQB103:02 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.
乳糜泻是一种小肠疾病，由遗传易感个体摄入麸质引发。乳糜泻与位于6号染色体上的人类白细胞抗原（HLA）密切相关，几乎100%的乳糜泻患者携带HLA - DQ2或HLA - DQ8异二聚体，其中大多数携带由DQA105:01/05:05、DQB102:01/02:02等位基因编码的HLA - DQ2，而其余患者携带由DQA103:01、DQB103:02等位基因编码的HLA - DQ8。在这项工作中，我们展示了一种36电极的多重电化学基因传感器阵列的开发，该阵列置于专用微流控平台内，共使用10种序列特异性探针进行快速中高分辨率HLA - DQ2/DQ8基因分型。利用目标序列和44个潜在干扰等位基因，包括单碱基错配区分，对设计探针的选择性进行了评估；结果表明具有良好的选择性。通过分析真实人类样本中HLA - DQ2/DQ8等位基因的存在情况，验证了电化学基因传感器阵列的性能，并与基于实验室的HLA分型结果进行比较，获得了极佳的相关性。
