Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS.
Identifieur interne : 005417 ( Main/Exploration ); précédent : 005416; suivant : 005418Cloning, sequencing, expression, and purification of SARS-associated coronavirus nucleocapsid protein for serodiagnosis of SARS.
Auteurs : Khalid Amine Timani [République populaire de Chine] ; Li Ye ; Linbai Ye ; Ying Zhu ; Zhenghui Wu ; Zuojiong GongSource :
- Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology [ 1386-6532 ] ; 2004.
Descripteurs français
- KwdFr :
- Analyse de séquence d'ADN, Anticorps antiviraux (sang), Clonage moléculaire, Données de séquences moléculaires, Escherichia coli (génétique), Escherichia coli (métabolisme), Humains, Protéines nucléocapside (), Protéines nucléocapside (génétique), Protéines nucléocapside (isolement et purification), Protéines nucléocapside (métabolisme), Protéines recombinantes (génétique), Protéines recombinantes (immunologie), Protéines recombinantes (métabolisme), Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Test ELISA, Tests sérologiques, Virus du SRAS (génétique), Virus du SRAS (immunologie), Virus du SRAS (métabolisme).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Escherichia coli, Protéines nucléocapside, Protéines recombinantes, Virus du SRAS.
- immunologie : Protéines recombinantes, Virus du SRAS.
- isolement et purification : Protéines nucléocapside.
- métabolisme : Escherichia coli, Protéines nucléocapside, Protéines recombinantes, Virus du SRAS.
- sang : Anticorps antiviraux.
- virologie : Syndrome respiratoire aigu sévère.
- Analyse de séquence d'ADN, Clonage moléculaire, Données de séquences moléculaires, Humains, Protéines nucléocapside, Test ELISA, Tests sérologiques.
English descriptors
- KwdEn :
- Antibodies, Viral (blood), Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Escherichia coli (genetics), Escherichia coli (metabolism), Humans, Molecular Sequence Data, Nucleocapsid Proteins (chemistry), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (isolation & purification), Nucleocapsid Proteins (metabolism), Recombinant Proteins (genetics), Recombinant Proteins (immunology), Recombinant Proteins (metabolism), SARS Virus (genetics), SARS Virus (immunology), SARS Virus (metabolism), Sequence Analysis, DNA, Serologic Tests, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology).
- MESH :
- chemical , blood : Antibodies, Viral.
- chemical , chemistry : Nucleocapsid Proteins.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Escherichia coli, Nucleocapsid Proteins, Recombinant Proteins, SARS Virus.
- chemical , immunology : Recombinant Proteins, SARS Virus.
- chemical , isolation & purification : Nucleocapsid Proteins.
- metabolism : Escherichia coli, Nucleocapsid Proteins, Recombinant Proteins, SARS Virus.
- virology : Severe Acute Respiratory Syndrome.
- Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Humans, Molecular Sequence Data, Sequence Analysis, DNA, Serologic Tests.
Abstract
A novel coronavirus has been associated with a worldwide outbreak of atypical pneumonia referred to as Severe Acute Respiratory Syndrome (SARS-CoV). SARS-CoV nucleocapsid (N) protein has been cloned sequenced and expressed in Escherichia coli strain. Purified N protein was used to measure the SARS-CoV specific IgG antibodies from 16 SARS-CoV infected patients' sera and from 131 control subjects using ELISA assay. Specific antibody responses to the purified recombinant N protein after 10, 20, and 30 days of disease onset were observed in 13 of 16 (81.3%), 16 of 16 (100%) and 16 of 16 (100%) SARS patients sera, respectively. Comparison of detection results with a commercially available diagnostic kit coated with a mixture of SARS-CoV viral proteins showed 9 of 16 (56.3%), 13 of 16 (81.3%), and 15 of 16 (93.7%) positive responses, respectively. None of 131 control sera gave positive reaction in either assay. This data suggests that the N protein of SARS-CoV is immunodominant and this ELISA based test assay for detecting the SARS-CoV N antigen may hold a significant value for SARS diagnosis.
DOI: 10.1016/j.jcv.2004.01.001
PubMed: 15163419
Affiliations:
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Le document en format XML
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sort="Gong, Zuojiong" uniqKey="Gong Z" first="Zuojiong" last="Gong">Zuojiong Gong</name></author></analytic><series><title level="j">Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology</title><idno type="ISSN">1386-6532</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (blood)</term><term>Cloning, Molecular</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Escherichia coli (genetics)</term><term>Escherichia coli (metabolism)</term><term>Humans</term><term>Molecular Sequence Data</term><term>Nucleocapsid Proteins (chemistry)</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (isolation & purification)</term><term>Nucleocapsid Proteins (metabolism)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (immunology)</term><term>Recombinant Proteins (metabolism)</term><term>SARS Virus (genetics)</term><term>SARS Virus (immunology)</term><term>SARS Virus (metabolism)</term><term>Sequence Analysis, DNA</term><term>Serologic Tests</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Severe Acute Respiratory Syndrome (virology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Analyse de séquence d'ADN</term><term>Anticorps antiviraux (sang)</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Escherichia coli (génétique)</term><term>Escherichia coli (métabolisme)</term><term>Humains</term><term>Protéines nucléocapside ()</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (isolement et purification)</term><term>Protéines nucléocapside (métabolisme)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (immunologie)</term><term>Protéines recombinantes (métabolisme)</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Test ELISA</term><term>Tests sérologiques</term><term>Virus du SRAS (génétique)</term><term>Virus du SRAS (immunologie)</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Nucleocapsid Proteins</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli</term><term>Nucleocapsid Proteins</term><term>Recombinant Proteins</term><term>SARS Virus</term></keywords><keywords 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coli</term><term>Protéines nucléocapside</term><term>Protéines recombinantes</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Humans</term><term>Molecular Sequence Data</term><term>Sequence Analysis, DNA</term><term>Serologic Tests</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de séquence d'ADN</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Protéines nucléocapside</term><term>Test ELISA</term><term>Tests sérologiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel coronavirus has been associated with a worldwide outbreak of atypical pneumonia referred to as Severe Acute Respiratory Syndrome (SARS-CoV). SARS-CoV nucleocapsid (N) protein has been cloned sequenced and expressed in Escherichia coli strain. Purified N protein was used to measure the SARS-CoV specific IgG antibodies from 16 SARS-CoV infected patients' sera and from 131 control subjects using ELISA assay. Specific antibody responses to the purified recombinant N protein after 10, 20, and 30 days of disease onset were observed in 13 of 16 (81.3%), 16 of 16 (100%) and 16 of 16 (100%) SARS patients sera, respectively. Comparison of detection results with a commercially available diagnostic kit coated with a mixture of SARS-CoV viral proteins showed 9 of 16 (56.3%), 13 of 16 (81.3%), and 15 of 16 (93.7%) positive responses, respectively. None of 131 control sera gave positive reaction in either assay. This data suggests that the N protein of SARS-CoV is immunodominant and this ELISA based test assay for detecting the SARS-CoV N antigen may hold a significant value for SARS diagnosis.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Hubei</li></region><settlement><li>Wuhan</li></settlement><orgName><li>Université de Wuhan</li></orgName></list><tree><noCountry><name sortKey="Gong, Zuojiong" sort="Gong, Zuojiong" uniqKey="Gong Z" first="Zuojiong" last="Gong">Zuojiong Gong</name><name sortKey="Wu, Zhenghui" sort="Wu, Zhenghui" uniqKey="Wu Z" first="Zhenghui" last="Wu">Zhenghui Wu</name><name sortKey="Ye, Li" sort="Ye, Li" uniqKey="Ye L" first="Li" last="Ye">Li Ye</name><name sortKey="Ye, Linbai" sort="Ye, Linbai" uniqKey="Ye L" first="Linbai" last="Ye">Linbai Ye</name><name sortKey="Zhu, Ying" sort="Zhu, Ying" uniqKey="Zhu Y" first="Ying" last="Zhu">Ying Zhu</name></noCountry><country name="République populaire de Chine"><region name="Hubei"><name sortKey="Timani, Khalid Amine" sort="Timani, Khalid Amine" uniqKey="Timani K" first="Khalid Amine" last="Timani">Khalid Amine Timani</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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