Methylation-specific electrochemical biosensing strategy for highly sensitive and quantitative analysis of promoter methylation of tumor-suppressor gene in real sample
Methylation-specific electrochemical biosensing strategy for highly sensitive and quantitative analysis of promoter methylation of tumor-suppressor gene in real sample
复制标题
甲基化特异性电化学生物传感策略，用于对真实样品中抑癌基因启动子甲基化进行高灵敏定量分析
DOI：
10.1016/j.jelechem.2016.03.001
复制
发表时间：
2016-07-15
影响因子：
4.5
通讯作者：
Zhao, Kebin
中科院分区：
文献类型：
--
作者：
Liu, Shenglin;Zhang, Xuejun;Zhao, Kebin
将亚硫酸氢盐转化和PCR扩增技术与高效电化学生物传感器相结合，建立了一种简便实用的甲基化特异性电化学生物传感策略。PCR和酶催化电化学传感的双重信号放大使该方法具有更高的灵敏度。设计了特异性引物和捕获探针，用于区分目的基因启动子区的CpG岛，进一步提高了特异性。所设计的方法对DNA甲基化分析显示出非常高的灵敏度和特异性，并且能够在10，000倍未甲基化的基因组DNA中响应约8个拷贝的分泌型卷曲相关蛋白2（SFRP 2）基因的甲基化启动子。该方法可用于乳腺癌组织中SFRP 2基因启动子甲基化的定量检测和去甲基化试剂处理后真实的乳腺癌组织中SFRP 2基因启动子甲基化状态的亚定量检测。以及监测对治疗剂的反应。(C)2016由Elsevier B.V.出版
A handy and practical methylation-specific electrochemical biosensing strategy was developed by integrating bisulfite conversion and PCR amplification with high efficient electrochemical biosensor. The dual signal amplification of PCR and enzyme-catalyzed electrochemical sensing resulted in higher sensitivity of the proposed method. The primers for PCR and capture probes for amplicons binding were specifically designed to distinguish different CpG islands in the promoter regions of target gene, which further enhanced the specificity. The designed method showed very high sensitivity and specificity for DNA methylation analysis and was able to respond to about 8 copies of methylated promotor of secreted frizzled-related protein 2 (SFRP2) gene in 10,000 times of unmethylated genomic DNA. Moreover, the established biosensor was successfully verified for quantitative detection of SFRP2 promotor methylation in breast primary tumor tissues and subtlemonitor of SFRP2 promotor methylation status changes in real sample with the demethylating agent treatment This proposed strategy presented a simple and pragmatic platform toward high sensitive and quantitative detection promoter methylation of tumor-suppressor gene for early diagnosis and prognostic assessment of cancer, as well as monitoring response to therapeutic agents. (C) 2016 Published by Elsevier B.V.