QDs-DNA nanosensor for the detection of hepatitis B virus DNA and the single-base mutants

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 th...

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Veröffentlicht in:Biosensors & bioelectronics 2010-04, Vol.25 (8), p.1934-1940
Hauptverfasser: Wang, Xiang, Lou, Xinhui, Wang, Yi, Guo, Qingchuan, Fang, Zheng, Zhong, Xinhua, Mao, Hongju, Jin, Qinghui, Wu, Lei, Zhao, Hui, Zhao, Jianlong
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container_end_page 1940
container_issue 8
container_start_page 1934
container_title Biosensors & bioelectronics
container_volume 25
creator Wang, Xiang
Lou, Xinhui
Wang, Yi
Guo, Qingchuan
Fang, Zheng
Zhong, Xinhua
Mao, Hongju
Jin, Qinghui
Wu, Lei
Zhao, Hui
Zhao, Jianlong
description 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 4.0 nM 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.
doi_str_mv 10.1016/j.bios.2010.01.007
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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 4.0 nM by using a multilabel counter. 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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 4.0 nM by using a multilabel counter. 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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 4.0 nM 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.</abstract><cop>Kidlington</cop><pub>Elsevier B.V</pub><pmid>20138498</pmid><doi>10.1016/j.bios.2010.01.007</doi><tpages>7</tpages></addata></record>
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subjects Biological and medical sciences
Biosensing Techniques - instrumentation
Biotechnology
DNA
DNA Mutational Analysis - instrumentation
DNA, Viral - analysis
DNA, Viral - genetics
Equipment Design
Equipment Failure Analysis
Fluorescence resonance energy transfer (FRET)
Fluorescence Resonance Energy Transfer - instrumentation
Fundamental and applied biological sciences. Psychology
Hepatitis B virus (HBV)
Hepatitis B virus - genetics
Mutation
Nanotechnology - instrumentation
Oligonucleotide Array Sequence Analysis - instrumentation
Quantum Dots
Quantum dots (QDs)
Reproducibility of Results
Sensitivity and Specificity
title QDs-DNA nanosensor for the detection of hepatitis B virus DNA and the single-base mutants
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