Nanowire-transistor based ultra-sensitive DNA methylation detection
Nanowire-transistor based ultra-sensitive DNA methylation detection
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DOI：
10.1016/j.bios.2007.08.017
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发表时间：
2008-01-18
影响因子：
12.6
通讯作者：
Maki, Gary K.
中科院分区：
文献类型：
--
作者：
Maki, Wusi C.;Mishra, Nirankar N.;Maki, Gary K.
在特定基因转录位点处的DNA甲基化的准确检测对于鉴定潜在的肿瘤形成是重要的，因为这种表观遗传改变可能导致保护免受肿瘤形成或修复受损DNA的肿瘤抑制基因的沉默。目前用于DNA甲基化检测的技术复杂且耗时。这项工作提出了第一个基于纳米线场效应晶体管（FET）的生物传感器技术，实现了简单和超灵敏的电子DNA甲基化检测，避免了复杂的亚硫酸氢盐处理和PCR扩增。p16（INK）基因是一种肿瘤抑制基因，其启动子是检测模型中的靶DNA。用磁珠捕获和浓缩靶DNA，并通过可逆结合过程释放到纳米FET的传感表面。甲基化的p16（INK）启动子被识别并结合到固定在纳米FET传感表面上的单克隆抗5-甲基胞嘧啶抗体。靶DNA分子的存在诱导了电子电荷，并改变了纳米晶体管的电子特性，从纳米晶体管产生可检测的电子信号。基于电子电荷的DNA甲基化检测方法简单、灵敏度高、成本低。检测灵敏度达到2.5 × 10（-19）mol，未观察到假阳性。(C)2007年由Elsevier B. V.出版。
Accurate detection of DNA methylation at specific gene transcription sites is important to identify potential tumor formation since this epigenetic alteration may result in silencing of tumor suppressor genes that protect against tumor formation or that repair damaged DNA. Current technologies used in DNA methylation detection are complicated and time consuming. This work presents the first nanowire field effect transistor (FET) based biosensor technology which achieves simple and ultra-sensitive electronic DNA methylation detection and avoids complicated bisulfite treatment and PCR amplification. The promoter of the p16(INK) gene, a tumor suppressor gene, is the target DNA in the detection model. The target DNA was captured and concentrated with magnetic beads, and released to the sensing surface of a nano-FET through a reversible binding process. The methylated p16(INK) promoter was recognized and bound to monoclonal anti-5-methylcytosine antibodies which were immobilized on the nano-FET sensing surface. The presence of the target DNA molecules induced electronic charge and changed the electronic properties of the nano-transistor from which detectable electronic signals are generated. The electronic charge based DNA methylation detection is simple and ultra-sensitive with the potential for low cost. The detection sensitivity was achieved at 2.5 x 10(-19) mol with no false positives observed. (C) 2007 Published by Elsevier B.V.