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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Wang, Yi Wang, Xiang Lou, Xinhui Fang, Zheng Zhao, Hui Guo, Qingchuan Zhong, Xinhua Jin, Qinghui Mao, Hongju Wu, Lei Zhao, Jianlong |
| Description | Author Affiliation: Wang X ( Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, PR China.) |
| Abstract | We report here a quantum dots-DNA (QDs-DNA) nanosensor based on fluorescence resonance energy transfer (FRET) for the detection of the target DNA and single mismatch in hepatitis B virus (HBV) gene. The proposed one-pot DNA detection method is simple, rapid and efficient due to the elimination of the washing and separation steps. In this study, the water-soluble CdSe/ZnS QDs were prepared by replacing the trioctylphosphine oxide (TOPO) on the surface of QDs with 3-mercaptopropionic acid (MPA). Subsequently, oligonucleotides were attached to the QDs surface to form functional QDs-DNA conjugates. Along with the addition of DNA targets and Cy5-modified signal DNAs into the QDs-DNA conjugates, sandwiched hybrids were formed. The resulting assembly brings the Cy5 fluorophore, the acceptor, and the QDs, the donor, into proximity, leading to fluorescence emission from the acceptor by means of FRET on illumination of the donor. In order to efficiently detect single-base mutants in HBV gene, oligonucleotide ligation assay was employed. If there existed a single-base mismatch, which could be recognized by the ligase, the detection probe was not ligated and no Cy5 emission was produced due to the lack of FRET. The feasibility of the proposed method was also demonstrated in the detection of synthetic 30-mer oliginucleotide targets derived from the HBV with a sensitivity of 4.0nM by using a multilabel counter. The method enables a simple and efficient detection that could be potentially used for high throughput and multiplex detections of target DNA and the mutants. |
| ISSN | 09565663 |
| Issue Number | 8 |
| Volume Number | 25 |
| e-ISSN | 18734235 |
| Journal | Biosensors and Bioelectronics |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2010-04-15 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Biosensing Techniques Instrumentation Dna Mutational Analysis Dna, Viral Analysis Fluorescence Resonance Energy Transfer Hepatitis B Virus Genetics Nanotechnology Quantum Dots Equipment Design Equipment Failure Analysis Oligonucleotide Array Sequence Analysis Reproducibility Of Results Sensitivity And Specificity Evaluation Studies Journal Article Research Support, Non-u.s. Gov't Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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