Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues

	Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues
	Zhu, Guichi; Liang, Li; Zhang, Chun-yang
刊名	ANALYTICAL CHEMISTRY
	2014
英文摘要	Sensitive and accurate analysis of microRNA (miRNA) expression is imperative for understanding the biological functions of miRNAs and the early diagnosis ofhuman cancers. Here, we develop a quencher-free fluorescent method for homogeneously sensitive detection of let-7a miRNA using the target-triggered recycling signal amplification in combination with a 2-aminopurine probe. The 2-aminopurine probe is characterized by the substitution of 2-aminopurine foradenine in the DNA strand and the quenching of 2-aminopurine fluorescence through its stacking interaction with the adjacent bases. The binding of target miRNA with the 2-aminopurme probe initiates the extension reaction in the presence of polymerase to produce the DNA duplexes. These DNA duplexes can be further cleaved by lambda exonuclease through the recycling digestion to release abundant free 2-arninopurines, leading to an enhanced fluorescence signal. The proposed method exhibits high sensitivity with a detection limit of 0.3 fmol, and it can even discriminate the single-base difference among the miRNA family members. More importantly, this method can accurately distinguish the expression of let-7a miRNA in human lung tissues between ten non small cell lung cancer (NSCLC) patients and ten healthy persons, holding a great potential for further application in early clinical diagnosis.
收录类别	SCI
原文出处	http://pubs.acs.org/doi/abs/10.1021/ac503365z
语种	英语
内容类型	期刊论文
源URL	[http://ir.siat.ac.cn:8080/handle/172644/6138]
专题	深圳先进技术研究院_医药所
作者单位	ANALYTICAL CHEMISTRY
推荐引用方式 GB/T 7714	Zhu, Guichi,Liang, Li,Zhang, Chun-yang. Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues[J]. ANALYTICAL CHEMISTRY,2014.
APA	Zhu, Guichi,Liang, Li,&Zhang, Chun-yang.(2014).Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues.ANALYTICAL CHEMISTRY.
MLA	Zhu, Guichi,et al."Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues".ANALYTICAL CHEMISTRY (2014).