High sensitive and direct fluorescence detection of single viral DNA sequences by integration of double strand probes onto microgels particles.
Identifieur interne : 001013 ( Main/Exploration ); précédent : 001012; suivant : 001014High sensitive and direct fluorescence detection of single viral DNA sequences by integration of double strand probes onto microgels particles.
Auteurs : A. Aliberti [Italie] ; A M Cusano [Italie] ; E. Battista [Italie] ; F. Causa [Italie] ; P A Netti [Italie]Source :
- The Analyst [ 1364-5528 ] ; 2016.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : DNA, Viral.
- chemical , chemistry : DNA, Viral, Fluorescent Dyes, Oligonucleotide Probes.
- chemical , genetics : DNA, Viral, Oligonucleotide Probes.
- Base Sequence, Gels, Limit of Detection, Spectrometry, Fluorescence.
Abstract
A novel class of probes for fluorescence detection was developed and combined to microgel particles for a high sensitive fluorescence detection of nucleic acids. A double strand probe with an optimized fluorescent-quencher couple was designed for the detection of different lengths of nucleic acids (39 nt and 100 nt). Such probe proved efficient in target detection in different contests and specific even in presence of serum proteins. The conjugation of double strand probes onto polymeric microgels allows for a sensitive detection of DNA sequences from HIV, HCV and SARS corona viruses with a LOD of 1.4 fM, 3.7 fM and 1.4 fM, respectively, and with a dynamic range of 10(-9)-10(-15) M. Such combination enhances the sensitivity of the detection of almost five orders of magnitude when compared to the only probe. The proposed platform based on the integration of innovative double strand probe into microgels particles represents an attractive alternative to conventional sensitive DNA detection technologies that rely on amplifications methods.
DOI: 10.1039/c5an02001h
PubMed: 26738973
Affiliations:
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Aliberti</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Cusano, A M" sort="Cusano, A M" uniqKey="Cusano A" first="A M" last="Cusano">A M Cusano</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Battista, E" sort="Battista, E" uniqKey="Battista E" first="E" last="Battista">E. Battista</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Causa, F" sort="Causa, F" uniqKey="Causa F" first="F" last="Causa">F. Causa</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Netti, P A" sort="Netti, P A" uniqKey="Netti P" first="P A" last="Netti">P A Netti</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2016">2016</date><idno type="RBID">pubmed:26738973</idno><idno type="pmid">26738973</idno><idno type="doi">10.1039/c5an02001h</idno><idno type="wicri:Area/PubMed/Corpus">000D23</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000D23</idno><idno type="wicri:Area/PubMed/Curation">000D23</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000D23</idno><idno type="wicri:Area/PubMed/Checkpoint">000C47</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000C47</idno><idno type="wicri:Area/Ncbi/Merge">002B78</idno><idno type="wicri:Area/Ncbi/Curation">002B78</idno><idno type="wicri:Area/Ncbi/Checkpoint">002B78</idno><idno type="wicri:Area/Main/Merge">001015</idno><idno type="wicri:Area/Main/Curation">001013</idno><idno type="wicri:Area/Main/Exploration">001013</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">High sensitive and direct fluorescence detection of single viral DNA sequences by integration of double strand probes onto microgels particles.</title><author><name sortKey="Aliberti, A" sort="Aliberti, A" uniqKey="Aliberti A" first="A" last="Aliberti">A. Aliberti</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Cusano, A M" sort="Cusano, A M" uniqKey="Cusano A" first="A M" last="Cusano">A M Cusano</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Battista, E" sort="Battista, E" uniqKey="Battista E" first="E" last="Battista">E. Battista</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Causa, F" sort="Causa, F" uniqKey="Causa F" first="F" last="Causa">F. Causa</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author><author><name sortKey="Netti, P A" sort="Netti, P A" uniqKey="Netti P" first="P A" last="Netti">P A Netti</name><affiliation wicri:level="1"><nlm:affiliation>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy.</nlm:affiliation><country xml:lang="fr">Italie</country><wicri:regionArea>Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy and Interdisciplinary Research Centre on Biomaterials (CRIB), University "Federico II", Piazzale Tecchio 80, 80125 Naples, Italy. paolo.netti@iit.it and Dipartimento di Ingegneria Chimica, dei Materiali e della Produzione Industriale (DICMAPI), University "Federico II", Piazzale Tecchio 80, 80125 Naples</wicri:regionArea><wicri:noRegion>80125 Naples</wicri:noRegion></affiliation></author></analytic><series><title level="j">The Analyst</title><idno type="eISSN">1364-5528</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Base Sequence</term><term>DNA, Viral (analysis)</term><term>DNA, Viral (chemistry)</term><term>DNA, Viral (genetics)</term><term>Fluorescent Dyes (chemistry)</term><term>Gels</term><term>Limit of Detection</term><term>Oligonucleotide Probes (chemistry)</term><term>Oligonucleotide Probes (genetics)</term><term>Spectrometry, Fluorescence</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN viral ()</term><term>ADN viral (analyse)</term><term>ADN viral (génétique)</term><term>Colorants fluorescents ()</term><term>Gels</term><term>Limite de détection</term><term>Sondes oligonucléotidiques ()</term><term>Sondes oligonucléotidiques (génétique)</term><term>Spectrométrie de fluorescence</term><term>Séquence nucléotidique</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA, Viral</term><term>Fluorescent Dyes</term><term>Oligonucleotide Probes</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>DNA, Viral</term><term>Oligonucleotide Probes</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN viral</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN viral</term><term>Sondes oligonucléotidiques</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Base Sequence</term><term>Gels</term><term>Limit of Detection</term><term>Spectrometry, Fluorescence</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN viral</term><term>Colorants fluorescents</term><term>Gels</term><term>Limite de détection</term><term>Sondes oligonucléotidiques</term><term>Spectrométrie de fluorescence</term><term>Séquence nucléotidique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel class of probes for fluorescence detection was developed and combined to microgel particles for a high sensitive fluorescence detection of nucleic acids. A double strand probe with an optimized fluorescent-quencher couple was designed for the detection of different lengths of nucleic acids (39 nt and 100 nt). Such probe proved efficient in target detection in different contests and specific even in presence of serum proteins. The conjugation of double strand probes onto polymeric microgels allows for a sensitive detection of DNA sequences from HIV, HCV and SARS corona viruses with a LOD of 1.4 fM, 3.7 fM and 1.4 fM, respectively, and with a dynamic range of 10(-9)-10(-15) M. Such combination enhances the sensitivity of the detection of almost five orders of magnitude when compared to the only probe. The proposed platform based on the integration of innovative double strand probe into microgels particles represents an attractive alternative to conventional sensitive DNA detection technologies that rely on amplifications methods. </div></front></TEI><affiliations><list><country><li>Italie</li></country></list><tree><country name="Italie"><noRegion><name sortKey="Aliberti, A" sort="Aliberti, A" uniqKey="Aliberti A" first="A" last="Aliberti">A. Aliberti</name></noRegion><name sortKey="Battista, E" sort="Battista, E" uniqKey="Battista E" first="E" last="Battista">E. Battista</name><name sortKey="Causa, F" sort="Causa, F" uniqKey="Causa F" first="F" last="Causa">F. Causa</name><name sortKey="Cusano, A M" sort="Cusano, A M" uniqKey="Cusano A" first="A M" last="Cusano">A M Cusano</name><name sortKey="Netti, P A" sort="Netti, P A" uniqKey="Netti P" first="P A" last="Netti">P A Netti</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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