Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions.

Non-cross-linking gold nanoparticle aggregation as a detection method for single-base substitutions.
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DOI:
10.1093/nar/gni007
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发表时间:
2005-01-07
影响因子:
14.9
通讯作者:
Maeda M
Maeda M
中科院分区:
生物学2区
文献类型:
--
作者:
Sato K;Hosokawa K;Maeda M

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DNA修饰的金纳米粒子在非交联构型中的聚集对表面结合的双链体的末端错配具有非凡的选择性。在本文中,我们证明了这种选择性检测单碱基置换的效用。通过标准方案制备样品:DNA提取、PCR扩增和单碱基引物延伸。寡核苷酸修饰的纳米颗粒正确地响应于来自引物延伸的未纯化产物:完全匹配的聚集和所有错配的分散。该方法对基因组DNA的适用性进行了测试与五个人类肿瘤细胞系,并验证了常规技术:质谱和直接测序。与现有的单碱基置换分析方法不同,该方法不需要专门的设备,为单核苷酸多态性的即时诊断开辟了新的可能性。
Aggregation of DNA-modified gold nanoparticles in a non-cross-linking configuration has extraordinary selectivity against terminal mismatch of the surface-bound duplex. In this paper, we demonstrate the utility of this selectivity for detection of single-base substitutions. The samples were prepared through standard protocols: DNA extraction, PCR amplification and single-base primer extension. Oligonucleotide-modified nanoparticles correctly responded to the unpurified products from the primer extension: aggregation for the full match and dispersion for all the mismatches. Applicability of this method to genomic DNA was tested with five human tumor cell lines, and verified by conventional technologies: mass spectrometry and direct sequencing. Unlike the existing methods for single-base substitution analysis, this method does not need specialized equipments, and opens up a new possibility of point-of-care diagnosis for single-nucleotide polymorphisms.
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发表时间: 1991-08-01
影响因子: 11.1
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