Department of Medicine, The University of Melbourne, Royal Melbourne Hospital, Victoria 3050, Australia; Department of Medical Biochemistry, Faculty of Medicine, Hasanuddin University, Makassar, South Sulawesi 90245, Indonesia.
Department of Medicine, The University of Melbourne, Royal Melbourne Hospital, Victoria 3050, Australia.
Biosens Bioelectron. 2018 Jul 15;111:174-183. doi: 10.1016/j.bios.2018.01.063. Epub 2018 Feb 2.
Prevention of life threatening hypersensitivity reactions to carbamazepine is possible through pre-treatment screening of the associated HLA-B15:02 risk allele. However, clinical implementation of screening is hindered by the high cost and slow turnaround of conventional HLA typing methods. We have developed an interdigitated electrode (IDE) biosensor platform utilizing loop mediated isothermal amplification (LAMP) that can rapidly detect the HLA-B15:02 allele. DNA amplification is followed by solid-phase hybridization of LAMP amplicons to a DNA probe immobilized on the IDE sensor surface, resulting in a change in sensor impedance. The testing platform does not require DNA extraction or post-amplification staining, achieving sample-to-answer in 1 h and 20 min. The platform was tested on 27 whole blood samples (14 HLA-B15:02 positive and 13 negative) with sensitivity of 92.9% and specificity of 84.6% when applying a cutoff of impedance change. Based on these characters the LAMP-IDE platform has potential to be further developed into point-of-care use to help overcome barriers in HLA-B15:02 screening.
通过对相关 HLA-B15:02 风险等位基因进行预处理筛查，可预防卡马西平引起的危及生命的过敏反应。然而，由于常规 HLA 分型方法成本高、周转时间长，限制了其临床应用。我们开发了一种基于交错电极（IDE）的生物传感器平台，利用环介导等温扩增（LAMP）技术，可以快速检测 HLA-B15:02 等位基因。在 DNA 扩增后，LAMP 扩增子与固定在 IDE 传感器表面的 DNA 探针进行固相杂交，导致传感器阻抗发生变化。该测试平台无需 DNA 提取或扩增后染色，可在 1 小时 20 分钟内实现从样本到结果的检测。该平台在 27 份全血样本（14 份 HLA-B15:02 阳性和 13 份阴性）上进行了测试，当应用阻抗变化的截止值时，其灵敏度为 92.9%，特异性为 84.6%。基于这些特性，LAMP-IDE 平台具有进一步开发为即时检测（POC）的潜力，以帮助克服 HLA-B15:02 筛查中的障碍。
