Fluorometric determination of the p53 cancer gene using strand displacement amplification on gold nanoparticles
Fluorometric determination of the p53 cancer gene using strand displacement amplification on gold nanoparticles
复制标题
使用金纳米颗粒上的链置换扩增荧光测定 p53 癌症基因
DOI：
10.1007/s00604-019-3609-0
复制
发表时间：
2019
影响因子：
5.7
通讯作者：
Yu Ruqin
中科院分区：
文献类型：
--
作者：
Zhao Weihua;Li Hongbo;Tang Yongqiong;Liu Mingbin;Wang Suqin;Yu Ruqin
描述了肿瘤抑制基因p53的荧光测定法。该方法基于金纳米颗粒（GNP）上的链置换扩增。使用FAM标记的发夹探针（HPP），其可以与GNP限制的接头链杂交，并且FAM标记的绿色荧光被GNP淬灭。在p53基因存在的情况下，它将与HPP杂交。这导致荧光恢复。然后引物与打开的HPP杂交并诱导聚合/置换反应。结果，杂交的p53基因被释放，并进而与GNP表面上的另一个HPP杂交。这引发了下一轮的杂交/酶促聚合/置换反应。这导致有效的链置换扩增并产生基本上放大的信号。该方法被称为GNP-HPP，因为它涉及使用GNP和HPP。该方法允许靶DNA（p53）定量至1.6 pM浓度，在5 pM至1 nM浓度范围内具有线性响应。此外，突变型p53基因可以容易地与野生型基因区分开。该方法是高度敏感的，选择性，并具有低的背景signal.Graphical abstractionSchematic介绍的链杂交信号放大系统（GNP-HPP）的基础上使用的金纳米粒子（GNP）作为淬灭源的肿瘤抑制基因p53的检测。将具有5′-末端修饰的荧光团的发夹探针（HPP）用作信号探针。
A fluorometric assay is described for the tumor suppressor gene p53. The method is based on strand displacement amplification on gold nanoparticles (GNPs). A FAM-labeled hairpin probe (HPP) is used that can hybridize to the GNP-confined linker strand, and the green fluorescence of the FAM label is quenched by the GNPs. In the presence of the p53 gene, it will hybridize with the HPP. This leads to fluorescence recovery. The primer then hybridizes with the opened HPP and induces the polymerization/displacement reactions. As a result, the hybridized p53 gene is released and, in turn, hybridizes with another HPP on the surface of the GNPs. This triggers the next round of hybridization/enzymatic polymerization/displacement reactions. This results in efficient strand displacement amplification and generates a substantially amplified signal. The method is referred to as GNP-HPP because it involves the use of GNPs and a HPP. The method allows the target DNA (p53) to be quantified down to 1.6 pM concentrations with a linear response in the 5 pM to 1 nM concentration range. In addition, mutant p53 genes can be easily distinguished from the wild-type gene. The method is highly sensitive, selective, and has a low background signal.Graphical abstractSchematic presentation of a chain hybridization signal amplification system (GNP-HPP) based on the use of gold nanoparticles (GNP) as a quenching source for the tumor suppressor gene p53 detection. The hairpin probe (HPP) having a 5′-end modified fluorophore was used as a signalling probe.