Detection of single-nucleotide polymorphisms using gold nanoparticles and single-strand-specific nucleases

被引:33
作者
Chen, Yen-Ting [1 ]
Hsu, Chiao-Ling [1 ]
Hou, Sao-Yi [1 ,2 ]
机构
[1] Natl Taipei Univ Technol, Inst Biotechnol, Taipei 106, Taiwan
[2] Natl Taipei Univ Technol, Dept Chem Engn, Taipei 106, Taiwan
关键词
single-nucleotide polymorphisms; gold nanoparticles; single-strand-specific nucleases;
D O I
10.1016/j.ab.2007.12.036
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The current study reports an assay approach that can detect single-nucleotide polymorphisms (SNPs) and identify the position of the point mutation through a single-strand-specific nuclease reaction and a gold nanoparticle assembly. The assay can be implemented via three steps: a single-strand-specific nuclease reaction that allows the enzyme to truncate the mutant DNA; a purification step that uses capture probe-gold nanoparticles and centrifugation; and a hybridization reaction that induces detector probe-gold nanoparticles, capture probe-gold nanoparticles, and the target DNA to form large DNA-linked three-dimensional aggregates of gold nanoparticles. At high temperature (63 degrees C in the current case), the purple color of the perfect match solution would not change to red, whereas a mismatched solution becomes red as the assembled gold nanoparticles separate. Using melting analysis, the position of the point mutation Could be identified. This assay provides a convenient colorimetric detection that enables point mutation identification without the need for expensive mass spectrometry. To our knowledge, this is the first report concerning SNP detection based on a single-strand-specific nuclease reaction and a gold nanoparticle assembly. (c) 2008 Published by Elsevier Inc.
引用
收藏
页码:299 / 305
页数:7
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