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Rolling circle amplification combined with gold nanoparticle aggregates for highly sensitive identification of single-nucleotide polymorphisms.

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WOS被引频次:129
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成果类型:
期刊论文
作者:
Jishan Li;Ting Deng;Xia Chu;Ronghua Yang;Jianhui Jiang;Guoli Shen;Ruqin Yu
通讯作者:
Chu, X.(xiachu@hnu.cn)
作者机构:
[Jishan Li; Ting Deng; Xia Chu; Jianhui Jiang; Ruqin Yu; Ronghua Yang; Guoli Shen] State Key Laboratory of Chem/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, 410082, China
[Ting Deng] Institute of Biological and Environmental Science and Technology, Central South University of Forestry and Technology, Changsha, 410004, China
通讯机构:
[Chu, Xia] Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chem Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China.
语种:
英文
关键词:
Assay methods - Colorimetric change - Colorimetric detection - DNA fragment - DNA ligases - DNA Probe - DNA targets - Endonucleases - Genomic DNA - Genotyping - Gold nanoparticle aggregates - Gold Nanoparticles - High fidelity - Highly sensitive - Naked-eye - Point mutations - Rolling circle amplifications - Single nucleotide polymorphisms - UV-vis spectroscopy - Wild types
期刊:
Analytical chemistry
ISSN:
0003-2700
年:
2010
卷:
82
期:
7
页码:
2811-2816
文献类别:
WOS:Article;EI:Journal article (JA)
所属学科:
ESI学科类别:化学;WOS学科类别:Chemistry, Analytical
入藏号:
WOS:000276004000031;EI:20101412831195;PMID:20192245
基金类别:
"973" National Key Basic Research Program [2007CB 310500]; National Natural Science Foundation of China [20775005, 20875027, 20975035, 07JJ1002]; Hunan University [521105668]
机构署名:
本校为其他机构
院系归属:
生命科学与技术学院
摘要:
A highly sensitive and specific colorimetry-based rolling circle amplification (RCA) assay method for single-nucleotide polymorphism genotyping has been developed. A circular template is generated by ligation upon the recognition of a point mutation on DNA targets. An RCA amplification is then initiated using the circular template in the presence of Phi29 polymerase. The RCA product can be digested by a restricting endonuclease, and the cleaved DNA fragments can mediate the aggregation of gold nanoparticle-tagged DNA probes. This causes a colorimetric change of the solution as the indicator of the mutation occurrence, which can be detected using UV-vis spectroscopy or viewed by naked eyes. On the basis of the high amplification efficiency of Phi29 polymerase, a mutated target of ∼70 fM can be detected in this assay. In addition, the protection of the circle template using phosphorothioated nucleotides allows the digestion reaction to be performed simultaneously in RCA. Moreover, DNA ligase offers high fidelity in distinguishing the mismatched bases at the ligation site, resulting in positive detection of mutant targets even when the ratio of the wildtype to the mutant is 10 000:1. The developed RCA-based colorimetric detection scheme was demonstrated for SNP typing of β-thalassemia gene at position -28 in genomic DNA. ©2010 American Chemical Society.
参考文献:
Baner J, 1998, NUCLEIC ACIDS RES, V26, P5073, DOI 10.1093/nar/26.22.5073
BARANY F, 1991, P NATL ACAD SCI USA, V88, P189, DOI 10.1073/pnas.88.1.189
Bjorheim J, 2005, ELECTROPHORESIS, V26, P2520, DOI 10.1002/elps.200410403
Blab GA, 2004, ANAL CHEM, V76, P495, DOI 10.1021/ac034987+
Carlton Victoria E. H., 2006, Human Genomics, V2, P391

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