文献类型：期刊文献

英文题名：An effective method based on real time fluorescence quenching for single nucleotide polymorphism detection

作者：Xu, Yichun[1,2];Han, Shuai[3];Huang, Xinhua[5];Zhuo, Shichao[6];Dai, Huiqing[5];Wang, Ke[4];Li, Zhou[3];Liu, Jianwen[1,2]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Sch Pharm, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, Shanghai Key Lab Chem Biol, Sch Pharm, Shanghai 200237, Peoples R China;[3]Southern Med Univ, Zhujiang Hosp, Dept Gen Surg, Guangzhou 510282, Guangdong, Peoples R China;[4]Shanghai Univ Tradit Chinese Med, Shuguang Hosp, Lab Integrat Med Surg, Shanghai 201203, Peoples R China;[5]Shanghai Aoyin Biotechnol Res & Dev Ltd Corp, Shanghai 201203, Peoples R China;[6]Cent Hosp Xuzhou, Dept Pathol, Xuzhou 221009, Peoples R China

年份：2014

卷号：186

起止页码：156

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20144400136433);WOS:【SCI-EXPANDED(收录号:WOS:000343836000019)】；

基金：This project was supported by Chuangxin Program (2012-AOYIN-001), the Fundamental Research Funds for the Central Universities, and the Natural Science Foundation of Guangdong Province (no. 9151051501000071).

语种：英文

外文关键词：Single nucleotide polymorphisms (SNPs); Fluorescence quenching; Sensitivity; PCR

摘要：In the Human Genome Project, the most common type of these variations is single nucleotide polymorphisms (SNPs). A large number of different SNP typing technologies have been developed in recent years. Enhancement and innovation for genotyping technologies are currently in progress. We described a rapid and effective method based on real time fluorescence quenching for SNP detection. The new method, Quenching-PCR, offering a single base extension method fully integrated with PCR which used a probe with quencher to eliminate fluorophor of the terminal base according to dideoxy sequencing method. In this platform, dideoxy sequencing reaction and obtaining values of real-time fluorescence occur simultaneously. The assay was validated by 106 DNA templates comparing with Sanger's sequencing and TaqMan assay. Compared with the results of DNA sequencing, the results of Quenching-PCR showed a high concordance rate of 93.40%, while the results of TaqMan platform showed a concordance rate of 92.45%, indicating that Quenching PCR and TaqMan assay were similar in accuracy. Therefore, Quenching PCR will be easily applicable and greatly accelerate the role of SNP detection in physiological processes of human health. (C) 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).