Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.
Identifieur interne : 002786 ( PubMed/Corpus ); précédent : 002785; suivant : 002787Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.
Auteurs : Alessandra Carattoli ; Paola Di Bonito ; Felicia Grasso ; Colomba Giorgi ; Francesco Blasi ; Matthias Niedrig ; Antonio CassoneSource :
- Journal of medical virology [ 0146-6615 ] ; 2005.
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
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pubmed:15834868Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.</title><author><name sortKey="Carattoli, Alessandra" sort="Carattoli, Alessandra" uniqKey="Carattoli A" first="Alessandra" last="Carattoli">Alessandra Carattoli</name><affiliation><nlm:affiliation>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy.</nlm:affiliation></affiliation></author><author><name sortKey="Di Bonito, Paola" sort="Di Bonito, Paola" uniqKey="Di Bonito P" first="Paola" last="Di Bonito">Paola Di Bonito</name></author><author><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name></author><author><name sortKey="Giorgi, Colomba" sort="Giorgi, Colomba" uniqKey="Giorgi C" first="Colomba" last="Giorgi">Colomba Giorgi</name></author><author><name sortKey="Blasi, Francesco" sort="Blasi, Francesco" uniqKey="Blasi F" first="Francesco" last="Blasi">Francesco Blasi</name></author><author><name sortKey="Niedrig, Matthias" sort="Niedrig, Matthias" uniqKey="Niedrig M" first="Matthias" last="Niedrig">Matthias Niedrig</name></author><author><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15834868</idno><idno type="pmid">15834868</idno><idno type="doi">10.1002/jmv.20338</idno><idno type="wicri:Area/PubMed/Corpus">002786</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002786</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.</title><author><name sortKey="Carattoli, Alessandra" sort="Carattoli, Alessandra" uniqKey="Carattoli A" first="Alessandra" last="Carattoli">Alessandra Carattoli</name><affiliation><nlm:affiliation>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy.</nlm:affiliation></affiliation></author><author><name sortKey="Di Bonito, Paola" sort="Di Bonito, Paola" uniqKey="Di Bonito P" first="Paola" last="Di Bonito">Paola Di Bonito</name></author><author><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name></author><author><name sortKey="Giorgi, Colomba" sort="Giorgi, Colomba" uniqKey="Giorgi C" first="Colomba" last="Giorgi">Colomba Giorgi</name></author><author><name sortKey="Blasi, Francesco" sort="Blasi, Francesco" uniqKey="Blasi F" first="Francesco" last="Blasi">Francesco Blasi</name></author><author><name sortKey="Niedrig, Matthias" sort="Niedrig, Matthias" uniqKey="Niedrig M" first="Matthias" last="Niedrig">Matthias Niedrig</name></author><author><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name></author></analytic><series><title level="j">Journal of medical virology</title><idno type="ISSN">0146-6615</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (blood)</term><term>Antigens, Viral (genetics)</term><term>Enzyme-Linked Immunosorbent Assay</term><term>Escherichia coli (genetics)</term><term>Escherichia coli (metabolism)</term><term>Humans</term><term>Immune Sera</term><term>Immunoglobulin G (blood)</term><term>Mice</term><term>Nucleocapsid Proteins (genetics)</term><term>Nucleocapsid Proteins (immunology)</term><term>Rabbits</term><term>Recombinant Proteins</term><term>SARS Virus (immunology)</term><term>Sensitivity and Specificity</term><term>Severe Acute Respiratory Syndrome (diagnosis)</term><term>Viral Matrix Proteins (genetics)</term><term>Viral Matrix Proteins (immunology)</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="genetics" xml:lang="en"><term>Escherichia coli</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" 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></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><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15834868</PMID><DateCompleted><Year>2005</Year><Month>09</Month><Day>16</Day></DateCompleted><DateRevised><Year>2019</Year><Month>12</Month><Day>10</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0146-6615</ISSN><JournalIssue CitedMedium="Print"><Volume>76</Volume><Issue>2</Issue><PubDate><Year>2005</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Journal of medical virology</Title><ISOAbbreviation>J. Med. Virol.</ISOAbbreviation></Journal><ArticleTitle>Recombinant protein-based ELISA and immuno-cytochemical assay for the diagnosis of SARS.</ArticleTitle><Pagination><MedlinePgn>137-42</MedlinePgn></Pagination><Abstract><AbstractText>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.</AbstractText><CopyrightInformation>Copyright 2005 Wiley-Liss, Inc.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Carattoli</LastName><ForeName>Alessandra</ForeName><Initials>A</Initials><AffiliationInfo><Affiliation>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità, Rome, Italy.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Di Bonito</LastName><ForeName>Paola</ForeName><Initials>P</Initials></Author><Author ValidYN="Y"><LastName>Grasso</LastName><ForeName>Felicia</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Giorgi</LastName><ForeName>Colomba</ForeName><Initials>C</Initials></Author><Author ValidYN="Y"><LastName>Blasi</LastName><ForeName>Francesco</ForeName><Initials>F</Initials></Author><Author ValidYN="Y"><LastName>Niedrig</LastName><ForeName>Matthias</ForeName><Initials>M</Initials></Author><Author ValidYN="Y"><LastName>Cassone</LastName><ForeName>Antonio</ForeName><Initials>A</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D023362">Evaluation Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J Med Virol</MedlineTA><NlmUniqueID>7705876</NlmUniqueID><ISSNLinking>0146-6615</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000914">Antibodies, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D000956">Antigens, Viral</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance 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