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Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues
Zhu, Guichi1; Liang, Li2; Zhang, Chun-yang1
刊名ANALYTICAL CHEMISTRY
2014-11-18
DOI10.1021/ac503365z
86期:22页:11410-11416
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]ROLLING-CIRCLE AMPLIFICATION ; ISOTHERMAL AMPLIFICATION ; GENE-EXPRESSION ; ULTRASENSITIVE DETECTION ; RECYCLING AMPLIFICATION ; REDUCED EXPRESSION ; MOLECULAR BEACONS ; CANCER DIAGNOSIS ; CHAIN-REACTION ; BREAST-CANCER
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 of human 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 for adenine 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.

语种英语
WOS记录号WOS:000345263300061
项目编号21325523 ; (2012) 433
资助机构National Natural Science Foundation of China ; Award for the Hundred Talent Program of the Chinese Academy of Science ; Fund for Shenzhen Engineering Laboratory of Single-molecule Detection and Instrument Development
引用统计
被引频次:35[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/68270
专题北京大学第三临床医学院
北京大学第三临床医学院_肿瘤治疗中心
作者单位1.Chinese Acad Sci, Shenzhen Inst Adv Technol, Single Mol Detect & Imaging Lab, Shenzhen 518055, Guangdong, Peoples R China
2.Peking Univ, Hosp 3, Dept Tumor Chemotherapy & Radiat Sickness, Beijing 100191, Peoples R China
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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,2014,86(22):11410-11416.
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,86(22),11410-11416.
MLA Zhu, Guichi,et al."Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues".ANALYTICAL CHEMISTRY 86.22(2014):11410-11416.
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