SrasV1, Main, Merge, bibRecord, 004B67

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 : 004B67 ( Main/Merge ); précédent : 004B66; suivant : 004B68

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 ; 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 Yuen

Source :

Journal of Clinical Microbiology [ 0095-1137 ] ; 2005.

RBID : PMC:1169156

Descripteurs français

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 (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.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1169156

DOI: 10.1128/JCM.43.7.3054-3058.2005
PubMed: 16000415
PubMed Central: 1169156

Links to Exploration step

PMC:1169156

Le document en format XML

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<term>Nucleocapsid Proteins</term>
<term>Peptides</term>
<term>Recombinant Proteins</term>
<term>Viral Envelope Proteins</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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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

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

Source :

Descripteurs français

English descriptors

Abstract

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