This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Giménez, L. G.
Right arrow Articles by Camacho, A. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Giménez, L. G.
Right arrow Articles by Camacho, A. G.

 Previous Article  |  Next Article 

Clinical and Vaccine Immunology, February 2009, p. 241-245, Vol. 16, No. 2
1071-412X/09/$08.00+0     doi:10.1128/CVI.00252-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

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{triangledown}

Luis G. Giménez, Jose Rojas, Almudena Rojas, Joaquín Mendoza, and Ana G. Camacho*

Laboratorios Vircell, SL, Granada, Spain

Received 11 July 2008/ Returned for modification 15 October 2008/ Accepted 17 November 2008

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.

* Corresponding author. Mailing address: Vircell, SL, C/Dominguez Ortiz, no. 1, Polígono Dos de Octubre, Santa Fe, 18320, Granada, Spain. Phone: 34 958 441264. Fax: 34 958 51012. E-mail: acamacho{at}vircell.com

{triangledown} Published ahead of print on 26 November 2008.

Clinical and Vaccine Immunology, February 2009, p. 241-245, Vol. 16, No. 2
1071-412X/09/$08.00+0     doi:10.1128/CVI.00252-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»