Colorimetric determination of BCR/ABL fusion genes using a nanocomposite consisting of Au@Pt nanoparticles covered with a PAMAM dendrimer and acting as a peroxidase mimic
Colorimetric determination of BCR/ABL fusion genes using a nanocomposite consisting of Au@Pt nanoparticles covered with a PAMAM dendrimer and acting as a peroxidase mimic
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使用由 Au@Pt 纳米颗粒组成的纳米复合材料比色测定 BCR/ABL 融合基因，纳米复合材料覆盖有 PAMAM 树枝状聚合物并充当过氧化物酶模拟物
DOI：
10.1007/s00604-018-2940-1
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发表时间：
2018-08-01
影响因子：
5.7
通讯作者：
Feng，Wenli
中科院分区：
文献类型：
--
作者：
Peng，Yang;Shen，Huawei;Feng，Wenli
本文介绍了一种检测BCR/ABL融合基因的比色法。将聚酰胺-胺（PAMAM）树枝状大分子置于过氧化物酶（POx）模拟Au@Pt纳米颗粒上，形成Au@Pt-PAMAM纳米复合材料。捕获DNA探针是设计的核酸链，其将靶DNA特异性结合到电极表面。捕获探针通过生物素和抗生物素蛋白相互作用连接到磁珠。捕获探针的发夹结构只能被互补的BCR/ABL DNA打开。这导致高度特异性的测定。Au@Pt-PAMAM的POx模拟特性导致通过H2O2和3，3，3 '，3'-四甲基联苯胺（TMB）反应形成蓝色染料，其通过酶标仪测量。在最佳条件下，BCR/ABL的吸光度在1 pM~100 nM浓度范围内呈线性增加，检测限低至190 fM。该方法选择性好，可用于真实的样品中融合基因的检测。可以想象，它在慢性粒细胞白血病患者的临床测试中具有很大的潜力。Au@PtNP是一种高效的催化剂，它与聚酰胺-胺（PAMAM）树枝状大分子结合，可以放大比色信号。通过引入链霉亲和素-磁珠去除非特异性信号，构建了一种新型的比色传感器，用于检测BCR/ABL融合基因。
A colorimetric assay is described for the detection of BCR/ABL fusion genes. Polyamidoamine (PAMAM) dendrimers were placed on peroxidase (POx) mimicking Au@Pt nanoparticles to form a nanocomposite of type Au@Pt-PAMAM. Capture DNA probe is a designed nucleic acid strand that specifically binds target DNA to the surface of the electrode. The capture probe was attached to magnetic beads via biotin and avidin interaction. The hairpin structure of the capture probe can only be opened by the complementary BCR/ABL DNA. This results in a highly specific assay. The POx-mimicking property of the Au@Pt-PAMAM causes the formation of a blue dye by reaction of H2O2and 3,3,3',3'-tetramethylbenzidine (TMB) which is measured by a microplate reader. Under optimum conditions, the absorbance increases linearly the 1 pM to 100 nM BCR/ABL concentration range, and the detection limit is as low as 190 fM. The method is highly selective and was successfully applied to the determination of fusion genes in spiked real samples. Conceivably, it possesses a large potential in clinical testing of patients suffering from chronic myeloid leukemia. Graphical abstract Au@PtNP, an efficient catalyst, is bound with polyamidoamine (PAMAM) dendrimer to amplify the colorimetric signal. With the introduction of streptavidin-magnetic beads to remove non-specific signals, a novel colorimetric sensor is constructed to detect BCR/ABL fusion genes.