Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia
Identifieur interne : 004E37 ( Main/Exploration ); précédent : 004E36; suivant : 004E38Differential sensitivities of severe acute respiratory syndrome (SARS) coronavirus spike polypeptide enzyme-linked immunosorbent assay (ELISA) and SARS coronavirus nucleocapsid protein ELISA for serodiagnosis of SARS coronavirus pneumonia
Auteurs : Patrick C. Y. Woo [Hong Kong] ; Susanna K. P. Lau [Hong Kong] ; Beatrice H. L. Wong [Hong Kong] ; Hoi-Wah Tsoi [Hong Kong] ; Ami M. Y. Fung [Hong Kong] ; Richard Y. T. Kao [Hong Kong] ; Kwok-Hung Chan [Hong Kong] ; J. S. Malik Peiris [Hong Kong] ; Kwok-Yung Yuen [Hong Kong]Source :
- Journal of clinical microbiology : (Print) [ 0095-1137 ] ; 2005.
Descripteurs français
- KwdFr :
- Anticorps antiviraux (sang), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (génétique), Glycoprotéines membranaires (immunologie), Humains, Peptides (immunologie), Protéines de l'enveloppe virale (génétique), Protéines de l'enveloppe virale (immunologie), Protéines nucléocapside (génétique), Protéines nucléocapside (immunologie), Protéines recombinantes (immunologie), Sensibilité et spécificité, Syndrome respiratoire aigu sévère (diagnostic), Syndrome respiratoire aigu sévère (virologie), Test ELISA, Tests sérologiques, Virus du SRAS (immunologie).
- MESH :
- diagnostic : Syndrome respiratoire aigu sévère.
- génétique : Glycoprotéines membranaires, Protéines de l'enveloppe virale, Protéines nucléocapside.
- immunologie : Glycoprotéines membranaires, Peptides, Protéines de l'enveloppe virale, Protéines nucléocapside, Protéines recombinantes, Virus du SRAS.
- sang : Anticorps antiviraux.
- virologie : Syndrome respiratoire aigu sévère.
- Pascal (Inist)
English descriptors
- KwdEn :
- Antibodies, Viral (blood), ELISA assay, Enzyme-Linked Immunosorbent Assay, Humans, Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Microbiology, Nucleocapsid, Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Peptides (immunology), Pneumonia, Polypeptide, Protein, Recombinant Proteins (immunology), SARS Virus (immunology), Sensitivity, Sensitivity and Specificity, Serologic Tests, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), Severe acute respiratory syndrome, Severe acute respiratory syndrome virus, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (genetics), Viral Envelope Proteins (immunology).
- MESH :
- chemical , blood : Antibodies, Viral.
- chemical , genetics : Membrane Glycoproteins, Nucleocapsid Proteins, Viral Envelope Proteins.
- chemical , immunology : Membrane Glycoproteins, Nucleocapsid Proteins, Peptides, Recombinant Proteins, Viral Envelope Proteins.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus.
- virology : Severe Acute Respiratory Syndrome.
- Enzyme-Linked Immunosorbent Assay, Humans, Sensitivity and Specificity, Serologic Tests, Spike Glycoprotein, Coronavirus.
Abstract
The use of recombinant severe acute respiratory syndrome-coronavirus (SARS-CoV) nucleocapsid protein (N) enzyme-linked immunosorbent assay (ELISA)-based antibody and antigen tests for diagnosis of SARS-CoV infections have been widely reported. However, no recombinant SARS-CoV spike protein (S)-based ELISA is currently available. In this article, we describe the problems and solutions of setting up the recombinant SARS-CoV S-based ELISA for antibody detection. The SARS-CoV S-based immunoglobulin M (IgM) and IgG ELISAs were evaluated and compared with the corresponding N-based ELISA for serodiagnosis of SARS-CoV pneumonia, using sera from 148 healthy blood donors who donated blood 3 years ago as controls and 95 SARS-CoV pneumonia patients in Hong Kong. Results obtained by the recombinant S (rS)-based IgG ELISA using the regenerated S prepared by dialysis with decreasing concentrations of urea or direct addition of different coating buffers, followed by addition of different regeneration buffer, identified 4 M urea and 1 M sarcosine for plate coating and no regeneration buffer as the most optimal conditions for antibody detection. The specificities of the S-based ELISA for IgG and IgM detection were 98.6% and 93.9%, with corresponding sensitivities of 58.9% and 74.7%, respectively. The sensitivity of the rN IgG ELISA (94.7%) is significantly higher than that of the rS IgG ELISA (P < 0.001), whereas the sensitivity of the rS IgM ELISA is significantly higher than that of the rN IgM ELISA (55.2%) (P < 0.01). An ELISA for detection of IgM against S and N could be more sensitive than one that detects IgM against N alone for serodiagnosis of SARS-CoV pneumonia.
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Le document en format XML
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<country>Hong Kong</country>
<wicri:noRegion>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
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<country>Hong Kong</country>
<wicri:noRegion>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
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</author>
<author><name sortKey="Chan, Kwok Hung" sort="Chan, Kwok Hung" uniqKey="Chan K" first="Kwok-Hung" last="Chan">Kwok-Hung Chan</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
<sZ>1 aut.</sZ>
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<country>Hong Kong</country>
<wicri:noRegion>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Malik Peiris, J S" sort="Malik Peiris, J S" uniqKey="Malik Peiris J" first="J. S." last="Malik Peiris">J. S. Malik Peiris</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
<sZ>1 aut.</sZ>
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<country>Hong Kong</country>
<wicri:noRegion>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>6 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>9 aut.</sZ>
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<country>Hong Kong</country>
<wicri:noRegion>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name>
<affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>3 aut.</sZ>
<sZ>4 aut.</sZ>
<sZ>5 aut.</sZ>
<sZ>6 aut.</sZ>
<sZ>7 aut.</sZ>
<sZ>8 aut.</sZ>
<sZ>9 aut.</sZ>
</inist:fA14>
<country>Hong Kong</country>
<wicri:noRegion>Department of Microbiology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
<affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</s1>
<s3>HKG</s3>
<sZ>1 aut.</sZ>
<sZ>2 aut.</sZ>
<sZ>6 aut.</sZ>
<sZ>8 aut.</sZ>
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<country>Hong Kong</country>
<wicri:noRegion>Research Centre of Infection and Immunology, Faculty of Medicine, The University of Hong Kong</wicri:noRegion>
</affiliation>
</author>
</analytic>
<series><title level="j" type="main">Journal of clinical microbiology : (Print)</title>
<title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title>
<idno type="ISSN">0095-1137</idno>
<imprint><date when="2005">2005</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><title level="j" type="main">Journal of clinical microbiology : (Print)</title>
<title level="j" type="abbreviated">J. clin. microbiol. : (Print)</title>
<idno type="ISSN">0095-1137</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibodies, Viral (blood)</term>
<term>ELISA assay</term>
<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Humans</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Microbiology</term>
<term>Nucleocapsid</term>
<term>Nucleocapsid Proteins (genetics)</term>
<term>Nucleocapsid Proteins (immunology)</term>
<term>Peptides (immunology)</term>
<term>Pneumonia</term>
<term>Polypeptide</term>
<term>Protein</term>
<term>Recombinant Proteins (immunology)</term>
<term>SARS Virus (immunology)</term>
<term>Sensitivity</term>
<term>Sensitivity and Specificity</term>
<term>Serologic Tests</term>
<term>Severe Acute Respiratory Syndrome (diagnosis)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Severe acute respiratory syndrome</term>
<term>Severe acute respiratory syndrome virus</term>
<term>Spike Glycoprotein, Coronavirus</term>
<term>Viral Envelope Proteins (genetics)</term>
<term>Viral Envelope Proteins (immunology)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Anticorps antiviraux (sang)</term>
<term>Glycoprotéine de spicule des coronavirus</term>
<term>Glycoprotéines membranaires (génétique)</term>
<term>Glycoprotéines membranaires (immunologie)</term>
<term>Humains</term>
<term>Peptides (immunologie)</term>
<term>Protéines de l'enveloppe virale (génétique)</term>
<term>Protéines de l'enveloppe virale (immunologie)</term>
<term>Protéines nucléocapside (génétique)</term>
<term>Protéines nucléocapside (immunologie)</term>
<term>Protéines recombinantes (immunologie)</term>
<term>Sensibilité et spécificité</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 (immunologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>Antibodies, Viral</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Nucleocapsid Proteins</term>
<term>Viral Envelope Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Membrane Glycoproteins</term>
<term>Nucleocapsid Proteins</term>
<term>Peptides</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope 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="génétique" xml:lang="fr"><term>Glycoprotéines membranaires</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Glycoprotéines membranaires</term>
<term>Peptides</term>
<term>Protéines de l'enveloppe virale</term>
<term>Protéines nucléocapside</term>
<term>Protéines recombinantes</term>
<term>Virus du SRAS</term>
</keywords>
<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</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>Enzyme-Linked Immunosorbent Assay</term>
<term>Humans</term>
<term>Sensitivity and Specificity</term>
<term>Serologic Tests</term>
<term>Spike Glycoprotein, Coronavirus</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Glycoprotéine de spicule des coronavirus</term>
<term>Humains</term>
<term>Sensibilité et spécificité</term>
<term>Test ELISA</term>
<term>Tests sérologiques</term>
<term>Virus syndrome respiratoire aigu sévère</term>
<term>Sensibilité</term>
<term>Polypeptide</term>
<term>Technique ELISA</term>
<term>Nucléocapside</term>
<term>Protéine</term>
<term>Microbiologie</term>
<term>Syndrome respiratoire aigu sévère</term>
<term>Pneumonie</term>
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<front><div type="abstract" xml:lang="en">The use of recombinant severe acute respiratory syndrome-coronavirus (SARS-CoV) nucleocapsid protein (N) enzyme-linked immunosorbent assay (ELISA)-based antibody and antigen tests for diagnosis of SARS-CoV infections have been widely reported. However, no recombinant SARS-CoV spike protein (S)-based ELISA is currently available. In this article, we describe the problems and solutions of setting up the recombinant SARS-CoV S-based ELISA for antibody detection. The SARS-CoV S-based immunoglobulin M (IgM) and IgG ELISAs were evaluated and compared with the corresponding N-based ELISA for serodiagnosis of SARS-CoV pneumonia, using sera from 148 healthy blood donors who donated blood 3 years ago as controls and 95 SARS-CoV pneumonia patients in Hong Kong. Results obtained by the recombinant S (rS)-based IgG ELISA using the regenerated S prepared by dialysis with decreasing concentrations of urea or direct addition of different coating buffers, followed by addition of different regeneration buffer, identified 4 M urea and 1 M sarcosine for plate coating and no regeneration buffer as the most optimal conditions for antibody detection. The specificities of the S-based ELISA for IgG and IgM detection were 98.6% and 93.9%, with corresponding sensitivities of 58.9% and 74.7%, respectively. The sensitivity of the rN IgG ELISA (94.7%) is significantly higher than that of the rS IgG ELISA (P < 0.001), whereas the sensitivity of the rS IgM ELISA is significantly higher than that of the rN IgM ELISA (55.2%) (P < 0.01). An ELISA for detection of IgM against S and N could be more sensitive than one that detects IgM against N alone for serodiagnosis of SARS-CoV pneumonia.</div>
</front>
</TEI>
<affiliations><list><country><li>Hong Kong</li>
</country>
</list>
<tree><country name="Hong Kong"><noRegion><name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C. Y." last="Woo">Patrick C. Y. Woo</name>
</noRegion>
<name sortKey="Chan, Kwok Hung" sort="Chan, Kwok Hung" uniqKey="Chan K" first="Kwok-Hung" last="Chan">Kwok-Hung Chan</name>
<name sortKey="Fung, Ami M Y" sort="Fung, Ami M Y" uniqKey="Fung A" first="Ami M. Y." last="Fung">Ami M. Y. Fung</name>
<name sortKey="Kao, Richard Y T" sort="Kao, Richard Y T" uniqKey="Kao R" first="Richard Y. T." last="Kao">Richard Y. T. Kao</name>
<name sortKey="Kao, Richard Y T" sort="Kao, Richard Y T" uniqKey="Kao R" first="Richard Y. T." last="Kao">Richard Y. T. Kao</name>
<name sortKey="Lau, Susanna K P" sort="Lau, Susanna K P" uniqKey="Lau S" first="Susanna K. P." last="Lau">Susanna K. P. Lau</name>
<name sortKey="Lau, Susanna K P" sort="Lau, Susanna K P" uniqKey="Lau S" first="Susanna K. P." last="Lau">Susanna K. P. Lau</name>
<name sortKey="Malik Peiris, J S" sort="Malik Peiris, J S" uniqKey="Malik Peiris J" first="J. S." last="Malik Peiris">J. S. Malik Peiris</name>
<name sortKey="Malik Peiris, J S" sort="Malik Peiris, J S" uniqKey="Malik Peiris J" first="J. S." last="Malik Peiris">J. S. Malik Peiris</name>
<name sortKey="Tsoi, Hoi Wah" sort="Tsoi, Hoi Wah" uniqKey="Tsoi H" first="Hoi-Wah" last="Tsoi">Hoi-Wah Tsoi</name>
<name sortKey="Wong, Beatrice H L" sort="Wong, Beatrice H L" uniqKey="Wong B" first="Beatrice H. L." last="Wong">Beatrice H. L. Wong</name>
<name sortKey="Woo, Patrick C Y" sort="Woo, Patrick C Y" uniqKey="Woo P" first="Patrick C. Y." last="Woo">Patrick C. Y. Woo</name>
<name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name>
<name sortKey="Yuen, Kwok Yung" sort="Yuen, Kwok Yung" uniqKey="Yuen K" first="Kwok-Yung" last="Yuen">Kwok-Yung Yuen</name>
</country>
</tree>
</affiliations>
</record>
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