Nucleic Acid Sandwich Hybridization Assay with Quantum Dot-Induced Fluorescence Resonance Energy Transfer for Pathogen Detection
Identifieur interne : 002230 ( Main/Exploration ); précédent : 002229; suivant : 002231Nucleic Acid Sandwich Hybridization Assay with Quantum Dot-Induced Fluorescence Resonance Energy Transfer for Pathogen Detection
Auteurs : Cheng-Chung Chou ; Yi-Han HuangSource :
- Sensors (Basel, Switzerland) [ 1424-8220 ] ; 2012.
Descripteurs français
- KwdFr :
- Animaux, Boîtes quantiques, Grippe chez les oiseaux (diagnostic), Grippe chez les oiseaux (virologie), Grippe humaine (diagnostic), Grippe humaine (virologie), Humains, Hybridation d'acides nucléiques, Hémagglutinines (), Oiseaux (virologie), Orthomyxoviridae (isolement et purification), Orthomyxoviridae (pathogénicité), Séquence nucléotidique, Transfert d'énergie par résonance de fluorescence.
- MESH :
- diagnostic : Grippe chez les oiseaux, Grippe humaine.
- isolement et purification : Orthomyxoviridae.
- pathogénicité : Orthomyxoviridae.
- virologie : Grippe chez les oiseaux, Grippe humaine, Oiseaux.
- Animaux, Boîtes quantiques, Humains, Hybridation d'acides nucléiques, Hémagglutinines, Séquence nucléotidique, Transfert d'énergie par résonance de fluorescence.
English descriptors
- KwdEn :
- Animals, Base Sequence, Birds (virology), Fluorescence Resonance Energy Transfer, Hemagglutinins (chemistry), Humans, Influenza in Birds (diagnosis), Influenza in Birds (virology), Influenza, Human (diagnosis), Influenza, Human (virology), Nucleic Acid Hybridization, Orthomyxoviridae (isolation & purification), Orthomyxoviridae (pathogenicity), Quantum Dots.
- MESH :
- chemical , chemistry : Hemagglutinins.
- diagnosis : Influenza in Birds, Influenza, Human.
- isolation & purification : Orthomyxoviridae.
- pathogenicity : Orthomyxoviridae.
- virology : Birds, Influenza in Birds, Influenza, Human.
- Animals, Base Sequence, Fluorescence Resonance Energy Transfer, Humans, Nucleic Acid Hybridization, Quantum Dots.
Abstract
This paper reports a nucleic acid sandwich hybridization assay with a quantum dot (QD)-induced fluorescence resonance energy transfer (FRET) reporter system. Two label-free hemagglutinin H5 sequences (60-mer DNA and 630-nt cDNA fragment) of avian influenza viruses were used as the targets in this work. Two oligonucleotides (16 mers and 18 mers) that specifically recognize two separate but neighboring regions of the H5 sequences were served as the capturing and reporter probes, respectively. The capturing probe was conjugated to QD655 (donor) in a molar ratio of 10:1 (probe-to-QD), and the reporter probe was labeled with Alexa Fluor 660 dye (acceptor) during synthesis. The sandwich hybridization assay was done in a 20 μL transparent, adhesive frame-confined microchamber on a disposable, temperature-adjustable indium tin oxide (ITO) glass slide. The FRET signal in response to the sandwich hybridization was monitored by a homemade optical sensor comprising a single 400 nm UV light-emitting diode (LED), optical fibers, and a miniature 16-bit spectrophotometer. The target with a concentration ranging from 0.5 nM to 1 μM was successfully correlated with both QD emission decrease at 653 nm and dye emission increase at 690 nm. To sum up, this work is beneficial for developing a portable QD-based nucleic acid sensor for on-site pathogen detection.
Url:
DOI: 10.3390/s121216660
PubMed: 23211753
PubMed Central: 3571803
Affiliations:
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Le document en format XML
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type="wicri:Area/Main/Exploration">002230</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Nucleic Acid Sandwich Hybridization Assay with Quantum Dot-Induced Fluorescence Resonance Energy Transfer for Pathogen Detection</title><author><name sortKey="Chou, Cheng Chung" sort="Chou, Cheng Chung" uniqKey="Chou C" first="Cheng-Chung" last="Chou">Cheng-Chung Chou</name></author><author><name sortKey="Huang, Yi Han" sort="Huang, Yi Han" uniqKey="Huang Y" first="Yi-Han" last="Huang">Yi-Han Huang</name></author></analytic><series><title level="j">Sensors (Basel, Switzerland)</title><idno type="eISSN">1424-8220</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Birds (virology)</term><term>Fluorescence Resonance Energy Transfer</term><term>Hemagglutinins (chemistry)</term><term>Humans</term><term>Influenza in Birds (diagnosis)</term><term>Influenza in Birds (virology)</term><term>Influenza, Human (diagnosis)</term><term>Influenza, Human (virology)</term><term>Nucleic Acid Hybridization</term><term>Orthomyxoviridae (isolation & purification)</term><term>Orthomyxoviridae (pathogenicity)</term><term>Quantum Dots</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Boîtes quantiques</term><term>Grippe chez les oiseaux (diagnostic)</term><term>Grippe chez les oiseaux (virologie)</term><term>Grippe humaine (diagnostic)</term><term>Grippe humaine (virologie)</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Hémagglutinines ()</term><term>Oiseaux (virologie)</term><term>Orthomyxoviridae (isolement et purification)</term><term>Orthomyxoviridae (pathogénicité)</term><term>Séquence nucléotidique</term><term>Transfert d'énergie par résonance de fluorescence</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Hemagglutinins</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Influenza in Birds</term><term>Influenza, Human</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Grippe chez les oiseaux</term><term>Grippe humaine</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Orthomyxoviridae</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Orthomyxoviridae</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>Orthomyxoviridae</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Orthomyxoviridae</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Grippe chez les oiseaux</term><term>Grippe humaine</term><term>Oiseaux</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Birds</term><term>Influenza in Birds</term><term>Influenza, Human</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Fluorescence Resonance Energy Transfer</term><term>Humans</term><term>Nucleic Acid Hybridization</term><term>Quantum Dots</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Boîtes quantiques</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Hémagglutinines</term><term>Séquence nucléotidique</term><term>Transfert d'énergie par résonance de fluorescence</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>This paper reports a nucleic acid sandwich hybridization assay with a quantum dot (QD)-induced fluorescence resonance energy transfer (FRET) reporter system. Two label-free hemagglutinin H5 sequences (60-mer DNA and 630-nt cDNA fragment) of avian influenza viruses were used as the targets in this work. Two oligonucleotides (16 mers and 18 mers) that specifically recognize two separate but neighboring regions of the H5 sequences were served as the capturing and reporter probes, respectively. The capturing probe was conjugated to QD655 (donor) in a molar ratio of 10:1 (probe-to-QD), and the reporter probe was labeled with Alexa Fluor 660 dye (acceptor) during synthesis. The sandwich hybridization assay was done in a 20 μL transparent, adhesive frame-confined microchamber on a disposable, temperature-adjustable indium tin oxide (ITO) glass slide. The FRET signal in response to the sandwich hybridization was monitored by a homemade optical sensor comprising a single 400 nm UV light-emitting diode (LED), optical fibers, and a miniature 16-bit spectrophotometer. The target with a concentration ranging from 0.5 nM to 1 μM was successfully correlated with both QD emission decrease at 653 nm and dye emission increase at 690 nm. To sum up, this work is beneficial for developing a portable QD-based nucleic acid sensor for on-site pathogen detection.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Alexander, D J" uniqKey="Alexander D">D.J. Alexander</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Beigel, J H" uniqKey="Beigel J">J.H. Beigel</name></author><author><name sortKey="Farrar, J" uniqKey="Farrar J">J. Farrar</name></author><author><name sortKey="Han, A M" uniqKey="Han A">A.M. Han</name></author><author><name sortKey="Hayden, F G" uniqKey="Hayden F">F.G. Hayden</name></author><author><name sortKey="Hyer, R" uniqKey="Hyer R">R. 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Daou</name></author><author><name sortKey="Li, L" uniqKey="Li L">L. Li</name></author><author><name sortKey="Reiss, P" uniqKey="Reiss P">P. Reiss</name></author><author><name sortKey="Josserand, V" uniqKey="Josserand V">V. Josserand</name></author><author><name sortKey="Texier, I" uniqKey="Texier I">I. Texier</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Algar, W R" uniqKey="Algar W">W.R. Algar</name></author><author><name sortKey="Krull, U J" uniqKey="Krull U">U.J. Krull</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list></list><tree><noCountry><name sortKey="Chou, Cheng Chung" sort="Chou, Cheng Chung" uniqKey="Chou C" first="Cheng-Chung" last="Chou">Cheng-Chung Chou</name><name sortKey="Huang, Yi Han" sort="Huang, Yi Han" uniqKey="Huang Y" first="Yi-Han" last="Huang">Yi-Han Huang</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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