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A Novel SNPs Detection Method Based on Gold Magnetic Nanoparticles Array and Single Base Extension

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WOS被引频次:199
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成果类型:
期刊论文
作者:
Li, Song;Liu, Hongna;Jia, Yingying;Deng, Yan;Zhang, Liming;Lu, Zhuoxuan;He, Nongyue
通讯作者:
He, NY
作者机构:
[Liu, Hongna; He, Nongyue; Jia, Yingying; Deng, Yan; Lu, Zhuoxuan; Zhang, Liming; Li, Song] Southeast Univ, State Key Lab Bioelect, Dept Biol Sci & Med Engn, Nanjing 210096, Jiangsu, Peoples R China.
[He, Nongyue; Deng, Yan; Lu, Zhuoxuan; Zhang, Liming; Li, Song] Hunan Univ Technol, Hunan Key Lab Green Packaging & Applicat Biol Nan, Zhuzhou 412007, Peoples R China.
通讯机构:
[He, NY] Southeast Univ, State Key Lab Bioelect, Dept Biol Sci & Med Engn, Sipailou 2, Nanjing 210096, Jiangsu, Peoples R China.
语种:
英文
关键词:
Magnetic beads;Dual-color SBE;Single nucleotide polymorphisms;Gold magnetic nanoparticles;Microarray
期刊:
THERANOSTICS
ISSN:
1838-7640
年:
2012
卷:
2
期:
10
页码:
967-975
文献类别:
WOS:Article
所属学科:
ESI学科类别:临床医学;WOS学科类别:Medicine, Research & Experimental
入藏号:
基金类别:
National Key Program for Developing Basic Research of China [2010CB933903]; NSFC [60927001, 60801007, 21205013, 61271056, 60971045]; Hunan Science and Technology Projects [2012SK3105, 2010sk2003]; Scientific Research Fund of Hunan Provincial Education Department [11A030]; China Postdoctoral Science Foundation [20100471362, 2012M511660]
机构署名:
本校为第一机构
院系归属:
包装与材料工程学院
摘要:
To fulfill the increasing need for large-scale genetic research, a high-throughput and automated SNPs genotyping method based on gold magnetic nanoparticles (GMNPs) array and dual-color single base extension has been designed. After amplification of DNA templates, biotinylated extension primers were captured by streptavidin coated gold magnetic nanoparticle (SA-GMNPs). Next a solid-phase, dual-color single base extension (SBE) reaction with the specific biotinylated primer was performed directly on the surface of the GMNPs. Finally, a "bead array" was fabricated by spotting GMNPs with fluorophore on a clean glass slide, and the genotype of each sample was discriminated by scanning the "bead array". MTHFR gene C677T polymorphism of 320 individual samples were interrogated using this method, the signal/noise ratio for homozygous samples were over 12.33, while the signal/noise ratio for heterozygous samples was near 1. Compared with other dual-color hybridization based genotyping methods, the method described here gives a higher signal/noise ratio and SNP loci can be identified with a high level of confidence. This assay has the advantage of eliminating the need for background subtraction and direct analysis of the fluorescence values of the GMNPs to determine their genotypes without the necessary procedures for purification and complex reduction of PCR products. The application of this strategy to large-scale SNP studies simplifies the process, and reduces the labor required to produce highly sensitive results while improving the potential for automation.
参考文献:
Komura D, 2006, GENOME RES, V16, P1575, DOI 10.1101/gr.5629106
Niu G, 2012, THERANOSTICS, V2, P413, DOI 10.7150/thno.3940
Naresh M, 2011, J BIOMED NANOTECHNOL, V7, P572, DOI 10.1166/jbn.2011.1323
Wang F, 2012, J BIOMED NANOTECHNOL, V8, P786, DOI 10.1166/jbn.2012.1446
Zhen ZP, 2012, THERANOSTICS, V2, P45, DOI 10.7150/thno.3448

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