RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein.
Identifieur interne : 002A43 ( Main/Exploration ); précédent : 002A42; suivant : 002A44RNA aptamer-based sensitive detection of SARS coronavirus nucleocapsid protein.
Auteurs : Dae-Gyun Ahn [Corée du Sud] ; Il-Ji Jeon ; Jung Dong Kim ; Min-Sun Song ; Seung-Ryul Han ; Seong-Wook Lee ; Hyungil Jung ; Jong-Won OhSource :
- The Analyst [ 1364-5528 ] ; 2009.
Descripteurs français
- KwdFr :
- MESH :
- analyse : Protéines nucléocapside.
- immunologie : Virus du SRAS.
- Analyse sur microréseau, Animaux, Aptamères nucléotidiques, Cellules Vero, Dosage immunologique, Mesures de luminescence.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Nucleocapsid Proteins.
- chemical : Aptamers, Nucleotide.
- immunology : SARS Virus.
- methods : Immunoassay.
- Animals, Chlorocebus aethiops, Luminescent Measurements, Microarray Analysis, Vero Cells.
Abstract
Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of a newly emerged disease SARS. The SARS-CoV nucleocapsid (N) protein is one of the most abundant structural proteins and serves as a diagnostic marker for accurate and sensitive detection of the virus. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant N protein, we selected a high-affinity RNA aptamer capable of binding to N protein with a dissociation constant of 1.65 nM. Electrophoretic mobility shift assays and RNA competition experiments showed that the selected aptamer recognized selectively the C-terminal region of N protein with high specificity. Using a chemiluminescence immunosorbent assay and a nanoarray aptamer chip with the selected aptamer as an antigen-capturing agent, we could sensitively detect N protein at a concentration as low as 2 pg/ml. These aptamer-antibody hybrid immunoassays may be useful for rapid, sensitive detection of SARS-CoV N protein.
DOI: 10.1039/b906788d
PubMed: 19684916
Affiliations:
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Lee</name></author><author><name sortKey="Jung, Hyungil" sort="Jung, Hyungil" uniqKey="Jung H" first="Hyungil" last="Jung">Hyungil Jung</name></author><author><name sortKey="Oh, Jong Won" sort="Oh, Jong Won" uniqKey="Oh J" first="Jong-Won" last="Oh">Jong-Won Oh</name></author></analytic><series><title level="j">The Analyst</title><idno type="eISSN">1364-5528</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Aptamers, Nucleotide</term><term>Chlorocebus aethiops</term><term>Immunoassay (methods)</term><term>Luminescent Measurements</term><term>Microarray Analysis</term><term>Nucleocapsid Proteins (analysis)</term><term>SARS Virus (immunology)</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Analyse sur microréseau</term><term>Animaux</term><term>Aptamères nucléotidiques</term><term>Cellules Vero</term><term>Dosage immunologique ()</term><term>Mesures de luminescence</term><term>Protéines nucléocapside (analyse)</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>Aptamers, Nucleotide</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Immunoassay</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Luminescent Measurements</term><term>Microarray Analysis</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse sur microréseau</term><term>Animaux</term><term>Aptamères nucléotidiques</term><term>Cellules Vero</term><term>Dosage immunologique</term><term>Mesures de luminescence</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of a newly emerged disease SARS. The SARS-CoV nucleocapsid (N) protein is one of the most abundant structural proteins and serves as a diagnostic marker for accurate and sensitive detection of the virus. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant N protein, we selected a high-affinity RNA aptamer capable of binding to N protein with a dissociation constant of 1.65 nM. Electrophoretic mobility shift assays and RNA competition experiments showed that the selected aptamer recognized selectively the C-terminal region of N protein with high specificity. Using a chemiluminescence immunosorbent assay and a nanoarray aptamer chip with the selected aptamer as an antigen-capturing agent, we could sensitively detect N protein at a concentration as low as 2 pg/ml. These aptamer-antibody hybrid immunoassays may be useful for rapid, sensitive detection of SARS-CoV N protein.</div></front></TEI><affiliations><list><country><li>Corée du Sud</li></country><region><li>Région capitale de Séoul</li></region><settlement><li>Séoul</li></settlement></list><tree><noCountry><name sortKey="Han, Seung Ryul" sort="Han, Seung Ryul" uniqKey="Han S" first="Seung-Ryul" last="Han">Seung-Ryul Han</name><name sortKey="Jeon, Il Ji" sort="Jeon, Il Ji" uniqKey="Jeon I" first="Il-Ji" last="Jeon">Il-Ji Jeon</name><name sortKey="Jung, Hyungil" sort="Jung, Hyungil" uniqKey="Jung H" first="Hyungil" last="Jung">Hyungil Jung</name><name sortKey="Kim, Jung Dong" sort="Kim, Jung Dong" uniqKey="Kim J" first="Jung Dong" last="Kim">Jung Dong Kim</name><name sortKey="Lee, Seong Wook" sort="Lee, Seong Wook" uniqKey="Lee S" first="Seong-Wook" last="Lee">Seong-Wook Lee</name><name sortKey="Oh, Jong Won" sort="Oh, Jong Won" uniqKey="Oh J" first="Jong-Won" last="Oh">Jong-Won Oh</name><name sortKey="Song, Min Sun" sort="Song, Min Sun" uniqKey="Song M" first="Min-Sun" last="Song">Min-Sun Song</name></noCountry><country name="Corée du Sud"><region name="Région capitale de Séoul"><name sortKey="Ahn, Dae Gyun" sort="Ahn, Dae Gyun" uniqKey="Ahn D" first="Dae-Gyun" last="Ahn">Dae-Gyun Ahn</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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