Qin Wei Jie, Yung Lin Yue Lanry
Department of Chemical and Biomolecular Engineering, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260.
Nucleic Acids Res. 2007;35(17):e111. doi: 10.1093/nar/gkm602. Epub 2007 Aug 24.
Sequence-specific DNA detection is important in various biomedical applications such as gene expression profiling, disease diagnosis and treatment, drug discovery and forensic analysis. Here we report a gold nanoparticle-based method that allows DNA detection and quantification and is capable of single nucleotide polymorphism (SNP) discrimination. The precise quantification of single-stranded DNA is due to the formation of defined nanoparticle-DNA conjugate groupings in the presence of target/linker DNA. Conjugate groupings were characterized and quantified by gel electrophoresis. A linear correlation between the amount of target DNA and conjugate groupings was found. For SNP detection, single base mismatch discrimination was achieved for both the end- and center-base mismatch. The method described here may be useful for the development of a simple and quantitative DNA detection assay.
序列特异性DNA检测在各种生物医学应用中都很重要,如基因表达谱分析、疾病诊断与治疗、药物发现和法医分析等。在此,我们报告了一种基于金纳米颗粒的方法,该方法可实现DNA检测和定量,并且能够区分单核苷酸多态性(SNP)。单链DNA的精确定量归因于在靶标/连接子DNA存在的情况下形成确定的纳米颗粒-DNA共轭聚集体。通过凝胶电泳对共轭聚集体进行表征和定量。发现靶标DNA的量与共轭聚集体之间存在线性相关性。对于SNP检测,无论是末端碱基错配还是中心碱基错配,都实现了单碱基错配区分。本文所述方法可能有助于开发一种简单的定量DNA检测方法。