Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education, Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University , Xi'an 710062, China.
ACS Appl Mater Interfaces. 2013 Nov 27;5(22):11741-6. doi: 10.1021/am4034119. Epub 2013 Nov 11.
Sensitive and specific detection of disease-related gene and single nucleotide polymorphism (SNP) is of great importance in cancer diagnosis. Here, a colorimetric and fluorescent approach is described for detection of the p53 gene and SNP in homogeneous solution by using gold nanorods (GNRs) as both colorimetric probe and fluorescence quencher. Hairpin oligonucleotide was utilized as DNA probe to ensure highly sequence-specific detection of target DNA. In the presence of target DNA, the formation of DNA duplex greatly changed the electrostatic interaction between GNR and DNAs, leading to an obvious change in fluorescence and colorimetric response. The detection limit of fluorescent and colorimetric assay is 0.26 pM and 0.3 nM, respectively. Both fluorescence and colorimetric strategies were able to effectively discriminate complementary DNA from single-base mismatched DNA, which is meaningful for cancer diagnosis. More important, target DNA can be detected as low as 10 nM by the naked eye. Furthermore, transmission electron microscopy and fluorescence anisotropy measurements demonstrated that the color change as well as fluorescence quenching is ascribed to the DNA hybridization-induced aggregation of GNRs. Therefore, the assay provided a fast, sensitive, cost-effective, and specific sensing platform for detecting disease-related gene and SNP.
灵敏且特异地检测疾病相关基因和单核苷酸多态性(SNP)对于癌症诊断具有重要意义。在这里,描述了一种在均相溶液中通过使用金纳米棒(GNRs)作为比色探针和荧光猝灭剂来检测 p53 基因和 SNP 的比色和荧光方法。发夹寡核苷酸被用作 DNA 探针,以确保对靶 DNA 进行高度序列特异性检测。在存在靶 DNA 的情况下,DNA 双链体的形成极大地改变了 GNR 与 DNA 之间的静电相互作用,导致荧光和比色响应的明显变化。荧光和比色测定的检测限分别为 0.26 pM 和 0.3 nM。荧光和比色两种策略都能够有效地区分互补 DNA 与单碱基错配 DNA,这对于癌症诊断具有重要意义。更重要的是,通过肉眼可以检测低至 10 nM 的靶 DNA。此外,透射电子显微镜和荧光各向异性测量表明,颜色变化和荧光猝灭归因于 DNA 杂交诱导的 GNR 聚集。因此,该测定法为检测疾病相关基因和 SNP 提供了一种快速、灵敏、具有成本效益且特异性的传感平台。