Recombinant protein‐based ELISA and immuno‐cytochemical assay for the diagnosis of SARS
Identifieur interne : 004B12 ( Main/Exploration ); précédent : 004B11; suivant : 004B13Recombinant protein‐based ELISA and immuno‐cytochemical assay for the diagnosis of SARS
Auteurs : Alessandra Carattoli [Italie] ; Paola Di Bonito [Italie] ; Felicia Grasso [Italie] ; Colomba Giorgi [Italie] ; Francesco Blasi [Italie] ; Matthias Niedrig [Allemagne] ; Antonio Cassone [Italie]Source :
- Journal of Medical Virology [ 0146-6615 ] ; 2005-06.
Descripteurs français
- KwdFr :
- Animaux, Anticorps antiviraux (sang), Antigènes viraux (génétique), Escherichia coli (génétique), Escherichia coli (métabolisme), Humains, Immunoglobuline G (sang), Lapins, Protéines de la matrice virale (génétique), Protéines de la matrice virale (immunologie), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), Protéines recombinantes, Sensibilité et spécificité, Souris, Syndrome respiratoire aigu sévère (diagnostic), Sérums immuns, Test ELISA, Virus du SRAS (immunologie).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Antigènes viraux, Escherichia coli, Protéines de la matrice virale, Protéines nucléocapside.
- immunologie : Protéines de la matrice virale, Protéines nucléocapside, Virus du SRAS.
- métabolisme : Escherichia coli.
- sang : Anticorps antiviraux, Immunoglobuline G.
- Animaux, Humains, Lapins, Protéines recombinantes, Sensibilité et spécificité, Souris, Sérums immuns, Test ELISA.
- Pascal (Inist)
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (blood), Antibody, Antigens, Viral (genetics), Coronavirus, Diagnosis, ELISA assay, Enzyme-Linked Immunosorbent Assay, Escherichia coli (genetics), Escherichia coli (metabolism), Humans, Immune Sera, Immunoglobulin G (blood), Mice, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Rabbits, Recombinant Proteins, Recombinant protein, SARS Virus (immunology), Sensitivity and Specificity, Serology, Severe Acute Respiratory Syndrome (diagnosis), Severe acute respiratory syndrome, Viral Matrix Proteins (genetics), Viral Matrix Proteins (immunology).
- MESH :
- chemical , blood : Antibodies, Viral, Immunoglobulin G.
- chemical , genetics : Antigens, Viral, Nucleocapsid Proteins, Viral Matrix Proteins.
- diagnosis : Severe Acute Respiratory Syndrome.
- genetics : Escherichia coli.
- chemical , immunology : Nucleocapsid Proteins, SARS Virus, Viral Matrix Proteins.
- metabolism : Escherichia coli.
- Animals, Enzyme-Linked Immunosorbent Assay, Humans, Immune Sera, Mice, Rabbits, Recombinant Proteins, Sensitivity and Specificity.
Abstract
A new Coronavirus (SARS‐CoV) is the aetiological agent of the severe acute respiratory syndrome (SARS). Because of the critical role played by serological assays for SARS diagnosis, an in‐house ELISA based on SARS‐CoV recombinant antigens was developed. The SARS‐CoV nucleocapsid protein (N), three N fragments (N1, N2, and N3) and the intraviral domain of the membrane protein (M2) were cloned and expressed in Escherichia coli as histidine‐tagged proteins. Six reference sera from SARS patients were used to detect virus‐specific IgG in an ELISA using each recombinant protein as coating antigen. High‐titre positive reactions were detected in all SARS positive sera. The specificity of the assay appears to be high as no positive reaction was detected in the sera of 20 healthy subjects and 73 patients with non‐SARS, low‐tract respiratory infections. Specific hyper‐immune sera to SARS‐CoV and the recombinant proteins, N, N1, N2, N3, and M2 were also generated in mice and rabbits. The specificity of these sera was confirmed by an immunocytochemical assay on biochips of SARS‐CoV infected and uninfected cells. J. Med. Virol. 76:137–142, 2005. © 2005 Wiley‐Liss, Inc.
Url:
DOI: 10.1002/jmv.20338
Affiliations:
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Le document en format XML
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nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Giorgi, Colomba" sort="Giorgi, Colomba" uniqKey="Giorgi C" first="Colomba" last="Giorgi">Colomba Giorgi</name><affiliation wicri:level="3"><country xml:lang="fr">Italie</country><wicri:regionArea>Department of Infectious, Parasitic and Immuno‐Mediated Diseases, Istituto Superiore di Sanità, Rome</wicri:regionArea><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Blasi, Francesco" sort="Blasi, Francesco" uniqKey="Blasi F" first="Francesco" last="Blasi">Francesco Blasi</name><affiliation wicri:level="3"><country xml:lang="fr">Italie</country><wicri:regionArea>Ospedale Maggiore Policlinico, University of Milan, Milan</wicri:regionArea><placeName><settlement type="city">Milan</settlement><region nuts="2">Lombardie</region></placeName></affiliation></author><author><name sortKey="Niedrig, Matthias" sort="Niedrig, 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wicri:curation="lc">Italie</country><wicri:regionArea>Correspondence address: Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome</wicri:regionArea><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Journal of Medical Virology</title><title level="j" type="alt">JOURNAL OF MEDICAL VIROLOGY</title><idno type="ISSN">0146-6615</idno><idno type="eISSN">1096-9071</idno><imprint><biblScope unit="vol">76</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="137">137</biblScope><biblScope unit="page" to="142">142</biblScope><biblScope unit="page-count">6</biblScope><publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher><pubPlace>Hoboken</pubPlace><date type="published" when="2005-06">2005-06</date></imprint><idno 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Matrix Proteins (genetics)</term><term>Viral Matrix Proteins (immunology)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (sang)</term><term>Antigènes viraux (génétique)</term><term>Escherichia coli (génétique)</term><term>Escherichia coli (métabolisme)</term><term>Humains</term><term>Immunoglobuline G (sang)</term><term>Lapins</term><term>Protéines de la matrice virale (génétique)</term><term>Protéines de la matrice virale (immunologie)</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (immunologie)</term><term>Protéines recombinantes</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Syndrome respiratoire aigu sévère (diagnostic)</term><term>Sérums immuns</term><term>Test ELISA</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term><term>Immunoglobulin G</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Antigens, Viral</term><term>Nucleocapsid Proteins</term><term>Viral Matrix 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></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Antigènes viraux</term><term>Escherichia coli</term><term>Protéines de la matrice virale</term><term>Protéines nucléocapside</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Protéines de la matrice virale</term><term>Protéines nucléocapside</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Nucleocapsid Proteins</term><term>SARS Virus</term><term>Viral Matrix Proteins</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Escherichia coli</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>Anticorps antiviraux</term><term>Immunoglobuline G</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Humans</term><term>Immune Sera</term><term>Mice</term><term>Rabbits</term><term>Recombinant Proteins</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Humains</term><term>Lapins</term><term>Protéines recombinantes</term><term>Sensibilité et spécificité</term><term>Souris</term><term>Sérums immuns</term><term>Test ELISA</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Anticorps</term><term>Coronavirus</term><term>Diagnostic</term><term>Protéine recombinante</term><term>Syndrome respiratoire aigu sévère</term><term>Sérologie</term><term>Technique ELISA</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A new Coronavirus (SARS‐CoV) is the aetiological agent of the severe acute respiratory syndrome (SARS). Because of the critical role played by serological assays for SARS diagnosis, an in‐house ELISA based on SARS‐CoV recombinant antigens was developed. The SARS‐CoV nucleocapsid protein (N), three N fragments (N1, N2, and N3) and the intraviral domain of the membrane protein (M2) were cloned and expressed in Escherichia coli as histidine‐tagged proteins. Six reference sera from SARS patients were used to detect virus‐specific IgG in an ELISA using each recombinant protein as coating antigen. High‐titre positive reactions were detected in all SARS positive sera. The specificity of the assay appears to be high as no positive reaction was detected in the sera of 20 healthy subjects and 73 patients with non‐SARS, low‐tract respiratory infections. Specific hyper‐immune sera to SARS‐CoV and the recombinant proteins, N, N1, N2, N3, and M2 were also generated in mice and rabbits. The specificity of these sera was confirmed by an immunocytochemical assay on biochips of SARS‐CoV infected and uninfected cells. J. Med. Virol. 76:137–142, 2005. © 2005 Wiley‐Liss, Inc.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Italie</li></country><region><li>Berlin</li><li>Latium</li><li>Lombardie</li></region><settlement><li>Berlin</li><li>Milan</li><li>Rome</li></settlement></list><tree><country name="Italie"><region name="Latium"><name sortKey="Carattoli, Alessandra" sort="Carattoli, Alessandra" uniqKey="Carattoli A" first="Alessandra" last="Carattoli">Alessandra Carattoli</name></region><name sortKey="Blasi, Francesco" sort="Blasi, Francesco" uniqKey="Blasi F" first="Francesco" last="Blasi">Francesco Blasi</name><name sortKey="Bonito, Paola Di" sort="Bonito, Paola Di" uniqKey="Bonito P" first="Paola Di" last="Bonito">Paola Di Bonito</name><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><name sortKey="Giorgi, Colomba" sort="Giorgi, Colomba" uniqKey="Giorgi C" first="Colomba" last="Giorgi">Colomba Giorgi</name><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name></country><country name="Allemagne"><region name="Berlin"><name sortKey="Niedrig, Matthias" sort="Niedrig, Matthias" uniqKey="Niedrig M" first="Matthias" last="Niedrig">Matthias Niedrig</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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