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学科主题: 临床医学
题名:
Quencher-Free Fluorescent Method for Homogeneously Sensitive Detection of MicroRNAs in Human Lung Tissues
作者: Zhu, Guichi1; Liang, Li2; Zhang, Chun-yang1
刊名: ANALYTICAL CHEMISTRY
发表日期: 2014-11-18
DOI: 10.1021/ac503365z
卷: 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
所属项目编号: 21325523 ; (2012) 433
项目资助者: 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
WOS记录号: WOS:000345263300061
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/68270
Appears in Collections:北京大学第三临床医学院_期刊论文

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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

Recommended Citation:
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.
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