Nonamplification Sandwich Assay Platform for Sensitive Nucleic Acid Detection Based on AuNPs Enumeration with the Dark-Field Microscope
Nonamplification Sandwich Assay Platform for Sensitive Nucleic Acid Detection Based on AuNPs Enumeration with the Dark-Field Microscope
复制标题
基于暗场显微镜金纳米粒子计数的非扩增夹心核酸检测灵敏检测平台
DOI：
10.1021/acs.analchem.6b00535
复制
发表时间：
2016-04-19
影响因子：
7.4
通讯作者：
Li, Na
中科院分区：
文献类型：
--
作者：
Li, Tian;Xu, Xiao;Li, Na
具有高灵敏度、方便实施以及试剂和仪器成本适中的简单有效的测定平台最适合于常规应用。基于本实验室建立的金纳米粒子（AuNP）计数信号读出模式，我们建立了一种非扩增夹心法检测阿尔茨海默病相关序列的核酸，检测限为3fM。这种基于AuNP计数的方法利用了暗场显微镜和磁性分离的独特且强的局部表面等离子体共振光散射。结果表明，存在20 nM随机DNA序列或质量高达靶标的10（6）倍的小牛胸腺DNA不会显著干扰测定信号。Hela细胞裂解物样品的加标回收率为109.3%（20 pM靶标）和110.5%（100 pM靶标），表明该方法在实际样品应用中的潜力。这种非扩增夹心测定平台原则上适用于其他测定，如免疫测定，因此将有望找到能够最好地利用简单性和灵敏度的广泛应用。
A simple and efficient assay platform with high sensitivity, convenient implementation, and moderate cost in reagents and instrumentation is most appropriate for routine applications. On the basis of the gold nanoparticle (AuNP) enumeration signal readout mode established in our laboratory, we have developed a nonamplification sandwich assay for nucleic acid detection with the 3 fM limit of detection for a sequence related to Alzheimer's disease. This AuNP counting based method takes advantages of the distinctive and strong localized surface plasmon resonance light scattering with the dark-field microscope and magnetic separation. It is shown that the presence of 20 nM random DNA sequence or calf thymus DNA with a mass up to 10(6)-fold of the targets do not significantly interfere with the assay signal. The spike recoveries of Hela cell lysate sample at 109.3% for 20 pM target and 110.5% for 100 pM target indicate the potential of this proposed method in practical sample applications. This nonamplification sandwich assay platform in principle is applicable to other assays such as the immunoassay and thus would be expected to find a breadth of applications that can make the best use of the simplicity and sensitivity.