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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 : 004E38

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

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 :

RBID : Pascal:05-0322075

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English descriptors

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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<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>
<sZ>9 aut.</sZ>
</inist:fA14>
<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>
<sZ>9 aut.</sZ>
</inist:fA14>
<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>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<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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