SrasV1, Ncbi, Curation, bibRecord, 001D96

Development of an Enzyme-Linked Immunosorbent Assay-Based Test with a Cocktail of Nucleocapsid and Spike Proteins for Detection of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific Antibody▿

Identifieur interne : 001D96 ( Ncbi/Curation ); précédent : 001D95; suivant : 001D97

Development of an Enzyme-Linked Immunosorbent Assay-Based Test with a Cocktail of Nucleocapsid and Spike Proteins for Detection of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific Antibody▿

Auteurs : Luis G. Giménez [Espagne] ; Jose Rojas [Espagne] ; Almudena Rojas [Espagne] ; Joaquín Mendoza [Espagne] ; Ana G. Camacho [Espagne]

Source :

Clinical and Vaccine Immunology : CVI [ 1556-6811 ] ; 2008.

RBID : PMC:2643541

Descripteurs français

English descriptors

KwdEn :
- Antibodies, Viral (blood), Antigens, Viral, Enzyme-Linked Immunosorbent Assay (methods), Humans, Immunoglobulin G (blood), Immunoglobulin M (blood), Membrane Glycoproteins (immunology), Nucleocapsid Proteins (immunology), Recombinant Proteins, SARS Virus (immunology), Sensitivity and Specificity, Severe Acute Respiratory Syndrome (diagnosis), Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (immunology).
MESH :
- chemical , blood : Antibodies, Viral, Immunoglobulin G, Immunoglobulin M.
- chemical , immunology : Membrane Glycoproteins, Nucleocapsid Proteins, Viral Envelope Proteins.
- chemical : Antigens, Viral, Recombinant Proteins, Spike Glycoprotein, Coronavirus.
- diagnosis : Severe Acute Respiratory Syndrome.
- immunology : SARS Virus.
- methods : Enzyme-Linked Immunosorbent Assay.
- Humans, Sensitivity and Specificity.

Abstract

A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2643541

DOI: 10.1128/CVI.00252-08
PubMed: 19038782
PubMed Central: 2643541

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PMC:2643541

Le document en format XML

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<term>Immunoglobulin G (blood)</term>
<term>Immunoglobulin M (blood)</term>
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<front><div type="abstract" xml:lang="en"><p>A new enzyme-linked immunosorbent assay (ELISA)-based immunoglobulin G (IgG)-plus-IgM antibody detection test for severe acute respiratory syndrome (SARS) has been developed by using a cocktail of four recombinant polypeptides as the antigen. These recombinant fragments were designed as parts of two different structural proteins from SARS-associated coronavirus (SARS-CoV). One recombinant polypeptide, S251-683, was designed as part of the spike glycoprotein, and the other three polypeptides comprised almost the whole nucleocapsid protein, avoiding the last 25 C-terminal amino acids. Immunization with a cocktail of these four polypeptides yielded a specific polyclonal antibody that is able to recognize SARS-CoV-infected cells by an immunofluorescence assay. This polypeptide cocktail was also used to set up an ELISA-based IgG-plus-IgM antibody detection test, which showed 99% specificity and 90% sensitivity upon evaluation using sera from 100 healthy negative controls and 20 SARS patients. Separate immunoreactivity assays with each recombinant polypeptide demonstrated that a combination of N and S protein fragments was more suitable than the individual peptides for developing a serological assay for SARS-CoV.</p>
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Development of an Enzyme-Linked Immunosorbent Assay-Based Test with a Cocktail of Nucleocapsid and Spike Proteins for Detection of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific Antibody▿

Development of an Enzyme-Linked Immunosorbent Assay-Based Test with a Cocktail of Nucleocapsid and Spike Proteins for Detection of Severe Acute Respiratory Syndrome-Associated Coronavirus-Specific Antibody▿

Source :

Descripteurs français

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Abstract

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