Recombinant protein-based elisa and immuno-cytochemical assay for the diagnosis of sars
Identifieur interne : 000588 ( PascalFrancis/Checkpoint ); précédent : 000587; suivant : 000589Recombinant 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.
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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.
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Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region 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 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INIST</idno><idno type="RBID">Pascal:05-0484475</idno><idno type="wicri:Area/PascalFrancis/Corpus">000582</idno><idno type="wicri:Area/PascalFrancis/Curation">000408</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000588</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000588</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">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 wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></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><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author><author><name sortKey="Grasso, Felicia" sort="Grasso, Felicia" uniqKey="Grasso F" first="Felicia" last="Grasso">Felicia Grasso</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region 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"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><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"><inist:fA14 i1="02"><s1>Ospedale Maggiore Policlinico, University of Milan</s1><s2>Milan</s2><s3>ITA</s3><sZ>5 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Milan</settlement><region nuts="2">Lombardie</region></placeName></affiliation></author><author><name sortKey="Niedrig, Matthias" sort="Niedrig, Matthias" uniqKey="Niedrig M" first="Matthias" last="Niedrig">Matthias Niedrig</name><affiliation wicri:level="3"><inist:fA14 i1="03"><s1>Robert Koch Institut</s1><s2>Berlin</s2><s3>DEU</s3><sZ>6 aut.</sZ></inist:fA14><country>Allemagne</country><placeName><region type="land" nuts="3">Berlin</region><settlement type="city">Berlin</settlement></placeName></affiliation></author><author><name sortKey="Cassone, Antonio" sort="Cassone, Antonio" uniqKey="Cassone A" first="Antonio" last="Cassone">Antonio Cassone</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></inist:fA14><country>Italie</country><placeName><settlement type="city">Rome</settlement><region nuts="2">Latium</region></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibody</term><term>Coronavirus</term><term>Diagnosis</term><term>ELISA assay</term><term>Recombinant protein</term><term>Serology</term><term>Severe acute respiratory syndrome</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Protéine recombinante</term><term>Technique ELISA</term><term>Diagnostic</term><term>Anticorps</term><term>Sérologie</term><term>Syndrome respiratoire aigu sévère</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><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0146-6615</s0></fA01><fA02 i1="01"><s0>JMVIDB</s0></fA02><fA03 i2="1"><s0>J. med. virol.</s0></fA03><fA05><s2>76</s2></fA05><fA06><s2>2</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Recombinant protein-based elisa and immuno-cytochemical assay for the diagnosis of sars</s1></fA08><fA11 i1="01" i2="1"><s1>CARATTOLI (Alessandra)</s1></fA11><fA11 i1="02" i2="1"><s1>DI BONITO (Paola)</s1></fA11><fA11 i1="03" i2="1"><s1>GRASSO (Felicia)</s1></fA11><fA11 i1="04" i2="1"><s1>GIORGI (Colomba)</s1></fA11><fA11 i1="05" i2="1"><s1>BLASI (Francesco)</s1></fA11><fA11 i1="06" i2="1"><s1>NIEDRIG (Matthias)</s1></fA11><fA11 i1="07" i2="1"><s1>CASSONE (Antonio)</s1></fA11><fA14 i1="01"><s1>Department of Infectious, Parasitic and Immuno-Mediated Diseases, Istituto Superiore di Sanità</s1><s2>Rome</s2><s3>ITA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>7 aut.</sZ></fA14><fA14 i1="02"><s1>Ospedale Maggiore Policlinico, University of Milan</s1><s2>Milan</s2><s3>ITA</s3><sZ>5 aut.</sZ></fA14><fA14 i1="03"><s1>Robert Koch Institut</s1><s2>Berlin</s2><s3>DEU</s3><sZ>6 aut.</sZ></fA14><fA20><s1>137-142</s1></fA20><fA21><s1>2005</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>17422</s2><s5>354000129811490010</s5></fA43><fA44><s0>0000</s0><s1>© 2005 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>17 ref.</s0></fA45><fA47 i1="01" i2="1"><s0>05-0484475</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Journal of medical virology</s0></fA64><fA66 i1="01"><s0>USA</s0></fA66><fC01 i1="01" l="ENG"><s0>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.</s0></fC01><fC02 i1="01" i2="X"><s0>002A05C10</s0></fC02><fC02 i1="02" i2="X"><s0>002B05C02J</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Protéine recombinante</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>Recombinant protein</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Proteína recombinante</s0><s5>05</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Technique ELISA</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>ELISA assay</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Técnica ELISA</s0><s5>06</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Diagnostic</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Diagnosis</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Diagnóstico</s0><s5>07</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Anticorps</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Antibody</s0><s5>08</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Anticuerpo</s0><s5>08</s5></fC03><fC03 i1="06" i2="X" l="FRE"><s0>Sérologie</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="ENG"><s0>Serology</s0><s5>09</s5></fC03><fC03 i1="06" i2="X" l="SPA"><s0>Serología</s0><s5>09</s5></fC03><fC03 i1="07" i2="X" l="FRE"><s0>Syndrome respiratoire aigu sévère</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="07" i2="X" l="ENG"><s0>Severe acute respiratory syndrome</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="07" i2="X" l="SPA"><s0>Síndrome respiratorio agudo severo</s0><s2>NM</s2><s5>14</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>Appareil respiratoire pathologie</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="ENG"><s0>Respiratory disease</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="SPA"><s0>Aparato respiratorio patología</s0><s5>13</s5></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Virose</s0></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Viral disease</s0></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Virosis</s0></fC07><fC07 i1="06" i2="X" l="FRE"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="ENG"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="SPA"><s0>Infección</s0></fC07><fC07 i1="07" i2="X" l="FRE"><s0>Poumon pathologie</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="ENG"><s0>Lung disease</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="SPA"><s0>Pulmón patología</s0><s5>16</s5></fC07><fN21><s1>339</s1></fN21></pA></standard></inist><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="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></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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