Department of Chemical and Biomolecular Engineering, Faculty of Engineering, National University of Singapore, Singapore 119260, Singapore.
Biosens Bioelectron. 2010 May 15;25(9):2021-5. doi: 10.1016/j.bios.2010.01.028. Epub 2010 Feb 1.
Nanoparticles are increasingly being used for applications in clinical diagnostics due to their unique physical and chemical properties. Gold nanoparticles, in particular, have unique optical properties allowing simplicity of detection methods. In this study, an assay based on dimeric assembly of gold nanoparticles was developed for discriminating single nucleotide mismatches. Only gel electrophoresis is needed for assay readout. No other sophisticated or expensive equipment is required. In addition, no false-positive was observed in the readout. We used this assay for genotyping mutations in the Duchenne muscular dystrophy (DMD) gene, the largest known in the human genome. Our results show that conjugating the gold nanoparticles with short DNA probes of 18 bases and 70 bases complimentary to target sequences allows specific discrimination between wild-type and mutant sequences for c.4150G > T (NM.004006.1) mutation in exon 30 of the DMD gene using a simple colorimetric detection. This method allows identification of both the patients as well as the carriers of the mutation who are at risk of transmitting the disease.
由于其独特的物理和化学性质，纳米粒子在临床诊断中的应用越来越广泛。特别是金纳米粒子具有独特的光学性质，使得检测方法变得简单。在这项研究中，我们开发了一种基于金纳米粒子二聚体组装的测定方法，用于区分单核苷酸错配。该测定方法的检测结果仅需凝胶电泳，不需要其他复杂或昂贵的设备。此外，在检测结果中没有观察到假阳性。我们使用该测定方法对杜氏肌营养不良症 (DMD) 基因中的基因突变进行基因分型，该基因是人类基因组中最大的已知基因。我们的结果表明，将金纳米粒子与互补于靶序列的 18 个碱基和 70 个碱基的短 DNA 探针缀合，允许使用简单的比色检测特异性区分野生型和突变型序列，用于 DMD 基因外显子 30 中的 c.4150G>T (NM.004006.1) 突变。该方法可识别患者以及突变的携带者，这些携带者有传播疾病的风险。
