Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.
Identifieur interne : 000E99 ( Ncbi/Checkpoint ); précédent : 000E98; suivant : 000F00Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.
Auteurs : Alessandra Carattoli [Italie] ; Paola Di Bonito ; Felicia Grasso ; Colomba Giorgi ; Francesco Blasi ; Matthias Niedrig ; Antonio CassoneSource :
- Journal of medical virology [ 0146-6615 ] ; 2005.
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.
English descriptors
- KwdEn :
- Animals, Antibodies, Viral (blood), Antigens, Viral (genetics), 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, SARS Virus (immunology), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), 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.
DOI: 10.1002/jmv.20338
PubMed: 15834868
Affiliations:
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pubmed:15834868Le document en format XML
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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.</div></front></TEI><affiliations><list><country><li>Italie</li></country><region><li>Latium</li></region><settlement><li>Rome</li></settlement></list><tree><noCountry><name sortKey="Blasi, Francesco" sort="Blasi, Francesco" uniqKey="Blasi F" first="Francesco" last="Blasi">Francesco Blasi</name><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><name sortKey="Di Bonito, Paola" sort="Di Bonito, Paola" uniqKey="Di Bonito P" first="Paola" last="Di Bonito">Paola Di Bonito</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><name sortKey="Niedrig, Matthias" sort="Niedrig, Matthias" uniqKey="Niedrig M" first="Matthias" last="Niedrig">Matthias Niedrig</name></noCountry><country name="Italie"><region name="Latium"><name sortKey="Carattoli, Alessandra" sort="Carattoli, Alessandra" uniqKey="Carattoli A" first="Alessandra" last="Carattoli">Alessandra Carattoli</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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