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基于金@铂纳米粒子的聚酰胺-胺树状大分子纳米复合材料模拟过氧化物酶用于 BCR/ABL 融合基因的比色测定

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.

机构信息

Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, No. 1 Yi Xue Yuan Road, Chongqing, 400016, People's Republic of China.

Traditional Chinese Medicine Hospital of Chongqing, Chongqing, 400021, People's Republic of China.

出版信息

Mikrochim Acta. 2018 Aug 4;185(8):401. doi: 10.1007/s00604-018-2940-1.

Abstract

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 HO and 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.

摘要

一种用于检测 BCR/ABL 融合基因的比色分析方法。聚酰胺胺(PAMAM)树枝状大分子被放置在过氧化物酶(POx)模拟金@铂纳米粒子上,形成 Au@Pt-PAMAM 纳米复合材料。捕获 DNA 探针是一种设计的核酸链,它特异性地将靶 DNA 结合到电极表面。捕获探针通过生物素和亲和素相互作用连接到磁珠上。捕获探针的发夹结构只能被互补的 BCR/ABL DNA 打开。这导致了高度特异性的测定。Au@Pt-PAMAM 的 POx 模拟特性导致 HO 和 3,3,3',3'-四甲基联苯胺(TMB)反应形成蓝色染料,这可以通过微孔板读数器测量。在最佳条件下,吸光度在 1 pM 至 100 nM 的 BCR/ABL 浓度范围内呈线性增加,检测限低至 190 fM。该方法具有高度选择性,并成功应用于掺入真实样本中融合基因的测定。可以想象,它在慢性髓性白血病患者的临床检测中具有很大的潜力。 图表摘要 作为一种高效的催化剂,金@铂纳米粒子与聚酰胺胺(PAMAM)树枝状大分子结合,放大比色信号。通过引入链霉亲和素-磁性珠来去除非特异性信号,构建了一种新型的比色传感器来检测 BCR/ABL 融合基因。

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