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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.
Clin Vaccine Immunol. 2009 Feb; 16(2):241-5.CV

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.

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Authors+Show Affiliations

Giménez LG
Laboratorios Vircell, SL, Granada, Spain.
Rojas J
No affiliation info available
Rojas A
No affiliation info available
Mendoza J
No affiliation info available
Camacho AG
No affiliation info available

MeSH

Antibodies, ViralAntigens, ViralCoronavirus Nucleocapsid ProteinsEnzyme-Linked Immunosorbent AssayHumansImmunoglobulin GImmunoglobulin MMembrane GlycoproteinsNucleocapsid ProteinsRecombinant ProteinsSARS VirusSensitivity and SpecificitySevere Acute Respiratory SyndromeSpike Glycoprotein, CoronavirusViral Envelope Proteins

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

19038782

Citation

Giménez, Luis G., et al. "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." Clinical and Vaccine Immunology : CVI, vol. 16, no. 2, 2009, pp. 241-5.
Giménez LG, Rojas J, Rojas A, et al. 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. Clin Vaccine Immunol. 2009;16(2):241-5.
Giménez, L. G., Rojas, J., Rojas, A., Mendoza, J., & Camacho, A. G. (2009). 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. Clinical and Vaccine Immunology : CVI, 16(2), 241-5. https://doi.org/10.1128/CVI.00252-08
Giménez LG, et al. 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. Clin Vaccine Immunol. 2009;16(2):241-5. PubMed PMID: 19038782.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Giménez,Luis G, AU - Rojas,Jose, AU - Rojas,Almudena, AU - Mendoza,Joaquín, AU - Camacho,Ana G, Y1 - 2008/11/26/ PY - 2008/11/29/pubmed PY - 2009/3/10/medline PY - 2008/11/29/entrez SP - 241 EP - 5 JF - Clinical and vaccine immunology : CVI JO - Clin Vaccine Immunol VL - 16 IS - 2 N2 - 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. SN - 1556-679X UR - https://www.unboundmedicine.com/medline/citation/19038782/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_ L2 - https://journals.asm.org/doi/10.1128/CVI.00252-08?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -
Grapherence [↓8 ↑23]

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