Gold-nanorod-based colorimetric and fluorescent approach for sensitive and specific assay of disease-related gene and mutation.
Gold-nanorod-based colorimetric and fluorescent approach for sensitive and specific assay of disease-related gene and mutation.
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DOI：
10.1021/am4034119
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发表时间：
2013-11
影响因子：
9.5
通讯作者：
Wenhong Wang;Yina Zhao;Yan Jin
中科院分区：
文献类型：
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作者：
Wenhong Wang;Yina Zhao;Yan Jin
敏感、特异地检测疾病相关基因和单核苷酸多态性（SNP）在癌症诊断中具有重要意义。在这里，描述了一种比色和荧光方法，用于通过使用金纳米棒（GNRs）作为比色探针和荧光猝灭剂来检测均相溶液中的p53基因和SNP。发夹寡核苷酸作为DNA探针，以确保高度序列特异性检测的目标DNA。在靶DNA存在下，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。荧光法和比色法的检测限分别为0.26 pM和0.3 nM。荧光法和比色法均能有效区分互补DNA和单碱基错配DNA，对癌症诊断有重要意义。更重要的是，靶DNA可以通过肉眼检测到低至10 nM。透射电子显微镜和荧光各向异性测量结果表明，颜色变化以及荧光猝灭归因于DNA杂交诱导的GNRs聚集。因此，该检测方法为检测疾病相关基因和SNP提供了一种快速、灵敏、经济、特异的传感平台。
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.