Adriaanse Marlou P M, Vreugdenhil Anita C E, Vastmans Véronique, Groeneveld Lisette, Molenbroeck Stefan, Schott Dina A, Voorter Christina E M, Tilanus Marcel G J
Department of Pediatrics & Nutrition and Toxicology Research Institute Maastricht (NUTRIM), Maastricht University Medical Center, Heerlen, the Netherlands.
Department of Transplantation Immunology and Tissue Typing, Maastricht University Medical Centre, Heerlen, the Netherlands.
J Gastroenterol Hepatol. 2016 Oct;31(10):1711-1716. doi: 10.1111/jgh.13331.
Human leukocyte antigen (HLA) typing is an important step in the diagnostic algorithm for celiac disease (CD) and is also used for screening purposes. Collection of blood is invasive and accompanied with emotional impact especially in children. Genetic technological progress now enables HLA typing from buccal cell samples. This study evaluated the reliability and feasibility of HLA typing for CD-associated HLA polymorphisms using buccal swabs as routine test in high-risk individuals.
Blood and buccal swabs of 77 children and adolescents with high risk for CD were prospectively collected in this cohort study. Buccal swab collection was performed either by the investigator at the outpatient clinic or by the patient or its parents at home. To evaluate the possibility of self-administration, three families performed the test at home. DNA was extracted using an adapted QIAamp method. Quantity, quality, and purity of DNA were recorded. HLA-DRB1, HLA-DQA1, and HLA-DQB1 typing was examined on buccal cell-derived and blood-derived DNA at low and, if necessary, high resolution level, using sequence-specific oligonucleotide and sequence-based typing, respectively.
DNA isolation using buccal swabs yielded a good quality and sufficient quantity of DNA to perform HLA-DQ typing in all individuals. HLA typing results on buccal cell-derived DNA were identical to typing on blood-derived DNA, also for the self-administered samples.
Introduction of the buccal swab test for HLA typing of CD risk in routine diagnostics can omit the current venipuncture and enables self-administration at home. Therefore, the buccal swab test is beneficial for individuals with a clinical suspicion for CD, as well as for screening purposes in high-risk populations.
人类白细胞抗原（HLA）分型是乳糜泻（CD）诊断流程中的重要步骤，也用于筛查。采血具有侵入性，且会带来情感影响，尤其是对儿童而言。基因技术的进步现在使得能够从颊细胞样本中进行HLA分型。本研究评估了使用颊拭子对与CD相关的HLA多态性进行HLA分型作为高危个体常规检测的可靠性和可行性。
在这项队列研究中前瞻性收集了77名CD高危儿童和青少年的血液及颊拭子。颊拭子采集由门诊的研究者进行，或由患者及其父母在家中进行。为评估自我采集的可能性，三个家庭在家中进行了检测。使用改良的QIAamp方法提取DNA。记录DNA的数量、质量和纯度。分别使用序列特异性寡核苷酸和基于序列的分型方法，在低分辨率水平（必要时在高分辨率水平）对颊细胞来源和血液来源的DNA进行HLA - DRB1、HLA - DQA1和HLA - DQB1分型检测。
使用颊拭子进行DNA分离产生了高质量且足够数量的DNA，可用于对所有个体进行HLA - DQ分型。颊细胞来源DNA的HLA分型结果与血液来源DNA的分型结果相同，自我采集的样本也是如此。
在常规诊断中引入用于CD风险HLA分型的颊拭子检测可以省去当前的静脉穿刺，并且能够在家中自我采集。因此，颊拭子检测对临床怀疑患有CD的个体以及高危人群的筛查均有益处。
