Mismatch Extension of DNA Polymerases and High-Accuracy Single Nucleotide Polymorphism Diagnostics by Gold Nanoparticle-Improved Isothermal Amplification
Chen, F; Zhao, Y; Fan, CH; Zhao, YX; Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China.
2015
发表期刊ANALYTICAL CHEMISTRY
ISSN0003-2700
卷号87期号:17页码:8718—8723
文章类型期刊文献
摘要Sequence Mismatches may induce nonspecific extension reaction, causing false results for SNP diagnostics. Herein, we. systematically investigated the impact of various 3'-terminal mismatches on isothermal amplification catalyzed by reptesentative DNA polymerases. Despite their diverse efficiencies depending on types of mismatch and kinds, of DNA: polymerase, all 12 kinds of single 3'-terminal mismatches induced the extension reaction. Generally, only several mismatches (primer-template, C-C, G-A, A-G, and A-A) present an observable inhibitory effect on the amplification reaction, whereas other mismatches trigger amplified signals as high as those of Watson-Crick pairs. The related mechanism was deeply discussed, and a primer-design guideline for specific SNP analysis was summarized. Furthermore, we found that the addition of appropriate gold nanoparticles (AuNPs) can significantly inhibit mismatch extension and enhance the amplification specificity. Also the high-accuracy SNP analysis of human blood genomic DNA has been demonstrated by AuNPs-improved isothermal amplification, the result of which was verified by sequencing (the gold standard method for SNP assay). Collectively, this work provides mechanistic insight into mismatch behavior and achieves accurate SNP diagnostics, holding great potential for the application in molecular diagnostics and personalized medicine.
关键词Rolling-circle Amplification Center-dot-t Nucleic-acids Base-pair Thermodynamics Sequences Nmr Quantification Replication Proteins
语种英语
WOS记录号WOS:000360773100020
引用统计
文献类型期刊论文
条目标识符http://ir.sinap.ac.cn/handle/331007/24994
专题中科院上海应用物理研究所2011-2018年
通讯作者Zhao, YX (reprint author), Xi An Jiao Tong Univ, Educ Minist, Sch Life Sci & Technol, Key Lab Biomed Informat Engn, Xianning West Rd, Xian 710049, Shaanxi, Peoples R China.
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Chen, F,Zhao, Y,Fan, CH,et al. Mismatch Extension of DNA Polymerases and High-Accuracy Single Nucleotide Polymorphism Diagnostics by Gold Nanoparticle-Improved Isothermal Amplification[J]. ANALYTICAL CHEMISTRY,2015,87(17):8718—8723.
APA Chen, F,Zhao, Y,Fan, CH,Zhao, YX,&Zhao, YX .(2015).Mismatch Extension of DNA Polymerases and High-Accuracy Single Nucleotide Polymorphism Diagnostics by Gold Nanoparticle-Improved Isothermal Amplification.ANALYTICAL CHEMISTRY,87(17),8718—8723.
MLA Chen, F,et al."Mismatch Extension of DNA Polymerases and High-Accuracy Single Nucleotide Polymorphism Diagnostics by Gold Nanoparticle-Improved Isothermal Amplification".ANALYTICAL CHEMISTRY 87.17(2015):8718—8723.
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