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基于 FeO@SiO@Au 功能化磁性纳米粒子的新型竞争化学发光 DNA 分析用于灵敏检测 p53 肿瘤抑制基因。

Novel Competitive Chemiluminescence DNA Assay Based on FeO@SiO@Au-Functionalized Magnetic Nanoparticles for Sensitive Detection of p53 Tumor Suppressor Gene.

机构信息

Electronic Materials Research Laboratory, Key Laboratory of the Ministry of Education & International Center for Dielectric Researh, Xi'an Jiaotong University, Xi'an, 710049, People's Republic of China.

Tongji University, Shanghai, People's Republic of China.

出版信息

Appl Biochem Biotechnol. 2019 Jan;187(1):152-162. doi: 10.1007/s12010-018-2808-1. Epub 2018 Jun 18.

Abstract

A simple, rapid response time and ultrahigh sensitive chemiluminescence (CL) DNA assay based on FeO@SiO@Au-functionalized magnetic nanoparticles (Au-MNPs) was developed for detection of p53 tumor suppressor gene. In this study, 2',6'-dimethylcarbonylphenyl-10-sulfopropyl acridinium-9-carboxylate 4'-NHS ester (NSP-DMAE-NHS), as a new kind of highly efficient luminescence reagent, was immobilized on the complementary sequence of the wild-type p53 (ssDNA) to improve the detection sensitivity. The optimal concentration of ssDNA-(NSP-DMAE-NHS) conjugates mixed with the wild-type p53 (wtp53) samples respectively. Then, the wtp53-Au-MNPs conjugates were added to continue the competitive reaction in the above solution. Subsequently, the Au-MNPs separated under magnetic field, measured by a homemade luminescent measurement system. Under optimal conditions, the method exhibited ultrasensitive sensitivity with a detection limit of 0.001 ng mL (0.16 pM), a wide range of liner response from 0.001 ng mL~6.6 μg mL. Therefore, the immunomagnetic nanocomposites-based detection strategy was rapid, low-cost, and highly sensitive that can be easily extended to the early diagnosis of cancer development and monitoring of patient therapy.

摘要

基于 FeO@SiO@Au 功能化磁性纳米粒子 (Au-MNPs) 的简单、快速响应时间和超高灵敏化学发光 (CL) DNA 测定法已被开发用于检测 p53 肿瘤抑制基因。在这项研究中,2',6'-二甲基羰基苯基-10-磺丙基吖啶-9-羧酸 4'-NHS 酯 (NSP-DMAE-NHS) 作为一种新型高效发光试剂被固定在野生型 p53 (ssDNA) 的互补序列上,以提高检测灵敏度。最佳浓度的 ssDNA-(NSP-DMAE-NHS) 与野生型 p53 (wtp53) 样品分别混合。然后,将 wtp53-Au-MNPs 缀合物加入上述溶液中继续进行竞争性反应。随后,在磁场下分离 Au-MNPs,并用自制的发光测量系统进行测量。在最佳条件下,该方法表现出超灵敏的灵敏度,检测限为 0.001ngmL(0.16pM),线性响应范围从 0.001ngmL 到 6.6μgmL。因此,基于免疫磁性纳米复合材料的检测策略快速、低成本且具有高灵敏度,可以很容易地扩展到癌症发展的早期诊断和患者治疗的监测。

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