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学科主题: 临床医学
题名:
A single quantum dot-based biosensor for DNA point mutation assay
作者: Tang, Wei1; Zhu, Guichi1; Liang, Li2; Zhang, Chun-Yang1,3
刊名: ANALYST
发表日期: 2015
DOI: 10.1039/c5an01270h
卷: 140, 期:17, 页:5936-5943
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]: CELL LUNG-CANCER ; FACTOR RECEPTOR MUTATIONS ; NUCLEOTIDE POLYMORPHISMS ; SENSITIVE METHOD ; EGFR ; NANOSENSOR
英文摘要:

Sensitive and selective detection of point mutation is essential to molecular biology research and early clinical diagnosis. Here, we demonstrate a single quantum dot (QD)-based biosensor for DNA point mutation assay. In this assay, a mutant target (G/C) remains unchanged after the endonuclease treatment, and the polymerase chain reaction (PCR) may be initiated with the assistance of primers and polymerase, generating a large number of mutant targets. The amplified mutant targets can be captured by biotinylated probes during the process of denaturation and annealing, and Cy5-dGTP may be assembled into the biotinylated probe with the catalysis of polymerase, leading to the formation of Cy5-labeled biotinylated probes. The Cy5-labeled biotinylated probes can be further assembled onto the QD surface to obtain a Cy5-DNA-QD complex, resulting in the generation of fluorescence resonance energy transfer (FRET) between the QD donor and the Cy5 receptor. The mutant targets can be quantitatively evaluated by the measurement of Cy5 counts by total internal reflection fluorescence (TIRF) microscopy. While in the presence of wild-type targets (T/A), no Cy5-dGTP can be assembled into the biotinylated probe due to the presence of a mismatch and consequently no FRET is observed. This single QD-based biosensor exhibits high sensitivity with a detection limit of 5.3 aM (or 32 copies) and can even discriminate as low as 0.01% variant frequency from the mixture of mutant targets and wild-type ones. Importantly, this biosensor can be used for genomic analysis in human lung cancer cells, and may be further applied for an early clinical diagnosis and personalized medicine.

语种: 英语
所属项目编号: 2011CB933600 ; 21325523
项目资助者: National Basic Research Program 973 ; National Natural Science Foundation of China ; Award for the Hundred Talent Program of Nanyue of Guangdong Province ; Award for Pengcheng Scholar of Shenzhen City
WOS记录号: WOS:000359217000017
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/51572
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, Peoples R China
2.Peking Univ, Hosp 3, Dept Tumor Chemotherapy & Radiat Sickness, Beijing 100191, Peoples R China
3.Shandong Normal Univ, Coll Chem Chem Engn & Mat Sci, Jinan 250014, Peoples R China

Recommended Citation:
Tang, Wei,Zhu, Guichi,Liang, Li,et al. A single quantum dot-based biosensor for DNA point mutation assay[J]. ANALYST,2015,140(17):5936-5943.
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