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 : 001042 ( Ncbi/Curation ); précédent : 001041; suivant : 001043Differential 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 ; Susanna K. P. Lau ; Beatrice H. L. Wong ; Hoi-Wah Tsoi ; Ami M. Y. Fung ; Richard Y. T. Kao ; Kwok-Hung Chan ; J. S. Malik Peiris ; Kwok-Yung YuenSource :
- Journal of Clinical Microbiology [ 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.
- Glycoprotéine de spicule des coronavirus, Humains, Sensibilité et spécificité, Test ELISA, Tests sérologiques.
English descriptors
- KwdEn :
- Antibodies, Viral (blood), Enzyme-Linked Immunosorbent Assay, Humans, Membrane Glycoproteins (genetics), Membrane Glycoproteins (immunology), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (immunology), Peptides (immunology), Recombinant Proteins (immunology), SARS Virus (immunology), Sensitivity and Specificity, Serologic Tests, Severe Acute Respiratory Syndrome (diagnosis), Severe Acute Respiratory Syndrome (virology), 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 (
Url:
DOI: 10.1128/JCM.43.7.3054-3058.2005
PubMed: 16000415
PubMed Central: 1169156
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PMC:1169156Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">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</title>
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<series><title level="j">Journal of Clinical Microbiology</title>
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<term>Enzyme-Linked Immunosorbent Assay</term>
<term>Humans</term>
<term>Membrane Glycoproteins (genetics)</term>
<term>Membrane Glycoproteins (immunology)</term>
<term>Nucleocapsid Proteins (genetics)</term>
<term>Nucleocapsid Proteins (immunology)</term>
<term>Peptides (immunology)</term>
<term>Recombinant Proteins (immunology)</term>
<term>SARS Virus (immunology)</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>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>
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<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>
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<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>
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<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term>
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<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term>
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<front><div type="abstract" xml:lang="en"><p>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 (<italic>P</italic>
< 0.001), whereas the sensitivity of the rS IgM ELISA is significantly higher than that of the rN IgM ELISA (55.2%) (<italic>P</italic>
< 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.</p>
</div>
</front>
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