Department of Pharmacology, Jinling Hospital, State Key Laboratory of Analytical Chemistry for Life Science, Medical School of Nanjing University, Nanjing, China.
Guangzhou Biotron Technology Co. Ltd., Guangzhou, China.
Clin Chem. 2017 Apr;63(4):852-860. doi: 10.1373/clinchem.2016.263996. Epub 2017 Feb 10.
Detecting DNA biomarkers related to personalized medicine could improve the outcome of drug therapy. However, personalized medicine in a resource-restrained hospital is very difficult because DNA biomarker detection should be performed by well-trained staff and requires expensive laboratory facilities.
We developed a gold nanoparticle-based "Tube-Lab" to enable DNA analysis in a closed tube. Gold nanoparticle-modified probes (GNPs) were used to construct an inexpensive and simple DNA sensor for signal readout. The method consists of 3 steps (template amplification, sequence identification, and GNP-based signal readout), bridged by an invasive reaction. With temperature control at each step, the 3 reactions proceed sequentially and automatically in a closed tube without any liquid transfer. We used Tube-Lab to detect different biomarkers in blood, tissue, and plasma, including US Food and Drug Administration-approved pharmacogenomic biomarkers (single nucleotide polymorphisms, somatic mutations).
The combination of PCR-based template replication and invader-based signal amplification allowed detection of approximately 6 copies of input DNA and the selective pick up 0.1% mutants from large amounts of background DNA. This method highly discriminated polymorphisms and somatic mutations from clinical samples and allowed a "liquid biopsy" assay with the naked eye.
Tube-Lab provides a promising and cost-effective approach for DNA biomarker analysis, including polymorphisms and somatic mutations from blood DNA, tissue DNA, or circulating tumor DNA in plasma, which are critical for personalized medicine.
检测与个性化医疗相关的 DNA 生物标志物可以改善药物治疗的效果。然而，资源有限的医院实施个性化医疗非常困难，因为 DNA 生物标志物检测应由经过良好培训的人员进行，并需要昂贵的实验室设备。
我们开发了一种基于金纳米颗粒的“管内实验室”（Tube-Lab），以实现封闭管内的 DNA 分析。金纳米颗粒修饰探针（GNPs）用于构建用于信号读出的廉价且简单的 DNA 传感器。该方法由 3 个步骤（模板扩增、序列识别和基于 GNP 的信号读出）组成，通过侵入性反应连接。通过在每个步骤进行温度控制，3 个反应在封闭管中顺序自动进行，无需任何液体转移。我们使用 Tube-Lab 来检测血液、组织和血浆中的不同生物标志物，包括美国食品和药物管理局批准的药物基因组生物标志物（单核苷酸多态性、体细胞突变）。
基于 PCR 的模板复制和基于入侵体的信号放大的组合允许检测约 6 个拷贝的输入 DNA，并且可以从大量背景 DNA 中选择性地提取 0.1%的突变体。该方法可以高度区分来自临床样本的多态性和体细胞突变，并允许进行肉眼可见的“液体活检”检测。
Tube-Lab 为 DNA 生物标志物分析提供了一种有前途且具有成本效益的方法，包括血液 DNA、组织 DNA 或血浆中循环肿瘤 DNA 的多态性和体细胞突变，这对个性化医疗至关重要。
