{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,4,15]],"date-time":"2025-04-15T04:55:01Z","timestamp":1744692901848},"reference-count":27,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2009,2,1]],"date-time":"2009-02-01T00:00:00Z","timestamp":1233446400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Clin Vaccine Immunol"],"published-print":{"date-parts":[[2009,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats: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.<\/jats:p>","DOI":"10.1128\/cvi.00252-08","type":"journal-article","created":{"date-parts":[[2008,11,27]],"date-time":"2008-11-27T02:47:39Z","timestamp":1227754059000},"page":"241-245","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":10,"title":["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"],"prefix":"10.1128","volume":"16","author":[{"given":"Luis G.","family":"Gime\u0301nez","sequence":"first","affiliation":[{"name":"Laboratorios Vircell, SL, Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jose","family":"Rojas","sequence":"additional","affiliation":[{"name":"Laboratorios Vircell, SL, Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Almudena","family":"Rojas","sequence":"additional","affiliation":[{"name":"Laboratorios Vircell, SL, Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joaqui\u0301n","family":"Mendoza","sequence":"additional","affiliation":[{"name":"Laboratorios Vircell, SL, Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ana G.","family":"Camacho","sequence":"additional","affiliation":[{"name":"Laboratorios Vircell, SL, Granada, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/0378-1119(89)90470-8","volume":"85","year":"1989","unstructured":"Brinkmann, U., R. 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