Room-Temperature Single-Nucleotide Polymorphism and Multiallele DNA Detection Using Fluorescent Nanocrystals and Microarrays
Identifieur interne : 000B18 ( Istex/Curation ); précédent : 000B17; suivant : 000B19Room-Temperature Single-Nucleotide Polymorphism and Multiallele DNA Detection Using Fluorescent Nanocrystals and Microarrays
Auteurs : Daniele Gerion [États-Unis] ; Fanqing Chen [États-Unis] ; Balaji Kannan [États-Unis] ; Aihua Fu [États-Unis] ; Wolfgang J. Parak [États-Unis] ; David J. Chen [États-Unis] ; Arunava Majumdar [États-Unis] ; A. Paul Alivisatos [États-Unis]Source :
- Analytical Chemistry [ 0003-2700 ] ; 2003.
Abstract
We report two cDNA microarray-based applications of DNA−nanocrystal conjugates, single-nucleotide polymorphism (SNP) and multiallele detections, using a commercial scanner and two sets of nanocrystals with orthogonal emissions. We focus on SNP mutation detection in the human p53 tumor suppressor gene, which has been found to be mutated in more than 50% of the known human cancers. DNA−nanocrystal conjugates are able to detect both SNP and single-base deletion at room temperature within minutes, with true-to-false signal ratios above 10. We also demonstrate microarray-based multiallele detection, using hybridization of multicolor nanocrystals conjugated to two sequences specific for the hepatitis B and hepatitis C virus, two common viral pathogens that inflict more than 10% of the population in the developing countries worldwide. The simultaneous detection of multiple genetic markers with microarrays and DNA−nanocrystal conjugates has no precedent and suggests the possibility of detecting an even greater number of bacterial or viral pathogens simultaneously.
Url:
DOI: 10.1021/ac034482j
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<sourceDesc><biblStruct><analytic><title level="a" type="main">Room-Temperature
Single-Nucleotide
Polymorphism and Multiallele DNA Detection Using
Fluorescent Nanocrystals and Microarrays</title>
<author><name sortKey="Gerion, Daniele" sort="Gerion, Daniele" uniqKey="Gerion D" first="Daniele" last="Gerion">Daniele Gerion</name>
<affiliation wicri:level="2"><mods:affiliation>Departmentof Chemistry and Department of Mechanical Engineering, Universityof California, Berkeley, California 94720,and Materials Sciences Division and Life Sciences Division, LawrenceBerkeley National Laboratory,Berkeley, California 94720</mods:affiliation>
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<affiliation><mods:affiliation> Materials Sciences Division, Lawrence Berkeley National Laboratory.</mods:affiliation>
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<affiliation wicri:level="2"><mods:affiliation> Present address: Lawrence Livermore National Laboratory,P.O. Box 808,L-415, Livermore, CA 94551.</mods:affiliation>
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<affiliation><mods:affiliation> To whom correspondence should be sent. E-mails: gerion1@llnl.gov(D.G.);f_chen@lbl.gov (F.C.).</mods:affiliation>
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<author><name sortKey="Chen, Fanqing" sort="Chen, Fanqing" uniqKey="Chen F" first="Fanqing" last="Chen">Fanqing Chen</name>
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<author><name sortKey="Kannan, Balaji" sort="Kannan, Balaji" uniqKey="Kannan B" first="Balaji" last="Kannan">Balaji Kannan</name>
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<country xml:lang="fr">États-Unis</country>
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<affiliation><mods:affiliation> Department of Mechanical Engineering, University of California.</mods:affiliation>
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<author><name sortKey="Fu, Aihua" sort="Fu, Aihua" uniqKey="Fu A" first="Aihua" last="Fu">Aihua Fu</name>
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<country xml:lang="fr">États-Unis</country>
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<affiliation><mods:affiliation> Department of Chemistry, University of California.</mods:affiliation>
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<author><name sortKey="Parak, Wolfgang J" sort="Parak, Wolfgang J" uniqKey="Parak W" first="Wolfgang J." last="Parak">Wolfgang J. Parak</name>
<affiliation wicri:level="2"><mods:affiliation>Departmentof Chemistry and Department of Mechanical Engineering, Universityof California, Berkeley, California 94720,and Materials Sciences Division and Life Sciences Division, LawrenceBerkeley National Laboratory,Berkeley, California 94720</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
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<affiliation><mods:affiliation> Department of Chemistry, University of California.</mods:affiliation>
</affiliation>
<affiliation><mods:affiliation> Materials Sciences Division, Lawrence Berkeley National Laboratory.</mods:affiliation>
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<author><name sortKey="Chen, David J" sort="Chen, David J" uniqKey="Chen D" first="David J." last="Chen">David J. Chen</name>
<affiliation wicri:level="2"><mods:affiliation>Departmentof Chemistry and Department of Mechanical Engineering, Universityof California, Berkeley, California 94720,and Materials Sciences Division and Life Sciences Division, LawrenceBerkeley National Laboratory,Berkeley, California 94720</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
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</affiliation>
<affiliation><mods:affiliation> Life Sciences Division, Lawrence Berkeley National Laboratory.</mods:affiliation>
</affiliation>
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<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Californie</region>
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</affiliation>
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<front><div type="abstract">We report two cDNA microarray-based applications of DNA−nanocrystal conjugates, single-nucleotide polymorphism (SNP) and multiallele detections, using a commercial scanner and two sets of nanocrystals with orthogonal emissions. We focus on SNP mutation detection in the human p53 tumor suppressor gene, which has been found to be mutated in more than 50% of the known human cancers. DNA−nanocrystal conjugates are able to detect both SNP and single-base deletion at room temperature within minutes, with true-to-false signal ratios above 10. We also demonstrate microarray-based multiallele detection, using hybridization of multicolor nanocrystals conjugated to two sequences specific for the hepatitis B and hepatitis C virus, two common viral pathogens that inflict more than 10% of the population in the developing countries worldwide. The simultaneous detection of multiple genetic markers with microarrays and DNA−nanocrystal conjugates has no precedent and suggests the possibility of detecting an even greater number of bacterial or viral pathogens simultaneously.</div>
</front>
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