Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.
Identifieur interne : 002074 ( Main/Exploration ); précédent : 002073; suivant : 002075Novel system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer.
Auteurs : Seong-Je Cho [Corée du Sud] ; Hye-Min Woo ; Ki-Sun Kim ; Jong-Won Oh ; Yong-Joo JeongSource :
- Journal of bioscience and bioengineering [ 1347-4421 ] ; 2011.
Descripteurs français
- KwdFr :
- ADN simple brin (), ADN simple brin (métabolisme), Alignement de séquences, Aptamères nucléotidiques (), Aptamères nucléotidiques (métabolisme), Données de séquences moléculaires, Humains, Protéines nucléocapside (génétique), Protéines nucléocapside (isolement et purification), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Séquence nucléotidique, Technique SELEX (), Technique de Western, Test ELISA, Virus du SRAS (génétique), Virus du SRAS (isolement et purification).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Protéines nucléocapside, Virus du SRAS.
- isolement et purification : Protéines nucléocapside, Virus du SRAS.
- métabolisme : ADN simple brin, Aptamères nucléotidiques.
- ADN simple brin, Alignement de séquences, Aptamères nucléotidiques, Données de séquences moléculaires, Humains, Sensibilité et spécificité, Séquence nucléotidique, Technique SELEX, Technique de Western, Test ELISA.
English descriptors
- KwdEn :
- Aptamers, Nucleotide (chemistry), Aptamers, Nucleotide (metabolism), Base Sequence, Blotting, Western, DNA, Single-Stranded (chemistry), DNA, Single-Stranded (metabolism), Enzyme-Linked Immunosorbent Assay, Humans, Molecular Sequence Data, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (isolation & purification), SARS Virus (genetics), SARS Virus (isolation & purification), SELEX Aptamer Technique (methods), Sensitivity and Specificity, Sequence Alignment, Severe Acute Respiratory Syndrome (diagnosis).
- MESH :
- chemical , chemistry : Aptamers, Nucleotide, DNA, Single-Stranded.
- chemical , genetics : Nucleocapsid Proteins.
- chemical , isolation & purification : Nucleocapsid Proteins.
- chemical , metabolism : Aptamers, Nucleotide, DNA, Single-Stranded.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- methods : SELEX Aptamer Technique.
- Base Sequence, Blotting, Western, Enzyme-Linked Immunosorbent Assay, Humans, Molecular Sequence Data, Sensitivity and Specificity, Sequence Alignment.
Abstract
The outbreak of severe acute respiratory syndrome (SARS) in 2002 affected thousands of people and an efficient diagnostic system is needed for accurate detection of SARS coronavirus (SARS CoV) to prevent or limit future outbreaks. Of the several SARS CoV structural proteins, the nucleocapsid protein has been shown to be a good diagnostic marker. In this study, an ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a K(d) of 4.93±0.30nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. Therefore, we believe that the selected ssDNA aptamer may be a good alternative detection probe for the rapid and sensitive detection of SARS.
DOI: 10.1016/j.jbiosc.2011.08.014
PubMed: 21920814
Affiliations:
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Le document en format XML
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Assay</term><term>Humans</term><term>Molecular Sequence Data</term><term>Sensitivity and Specificity</term><term>Sequence Alignment</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN simple brin</term><term>Alignement de séquences</term><term>Aptamères nucléotidiques</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Sensibilité et spécificité</term><term>Séquence nucléotidique</term><term>Technique SELEX</term><term>Technique de Western</term><term>Test ELISA</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The outbreak of severe acute respiratory syndrome (SARS) in 2002 affected thousands of people and an efficient diagnostic system is needed for accurate detection of SARS coronavirus (SARS CoV) to prevent or limit future outbreaks. Of the several SARS CoV structural proteins, the nucleocapsid protein has been shown to be a good diagnostic marker. In this study, an ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a K(d) of 4.93±0.30nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. Therefore, we believe that the selected ssDNA aptamer may be a good alternative detection probe for the rapid and sensitive detection of SARS.</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="Jeong, Yong Joo" sort="Jeong, Yong Joo" uniqKey="Jeong Y" first="Yong-Joo" last="Jeong">Yong-Joo Jeong</name><name sortKey="Kim, Ki Sun" sort="Kim, Ki Sun" uniqKey="Kim K" first="Ki-Sun" last="Kim">Ki-Sun Kim</name><name sortKey="Oh, Jong Won" sort="Oh, Jong Won" uniqKey="Oh J" first="Jong-Won" last="Oh">Jong-Won Oh</name><name sortKey="Woo, Hye Min" sort="Woo, Hye Min" uniqKey="Woo H" first="Hye-Min" last="Woo">Hye-Min Woo</name></noCountry><country name="Corée du Sud"><region name="Région capitale de Séoul"><name sortKey="Cho, Seong Je" sort="Cho, Seong Je" uniqKey="Cho S" first="Seong-Je" last="Cho">Seong-Je Cho</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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