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Article: 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
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TitleDifferential 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
 
AuthorsWoo, PCY1 1
Lau, SKP1 1
Wong, BHL1
Tsoi, HW1
Fung, AMY1
Kao, RYT1 1
Chan, KH1
Peiris, JSM1 1
Yuen, KY1 1 1
 
KeywordsAntibodies, Viral - blood
Membrane Glycoproteins - genetics - immunology
Nucleocapsid Proteins - genetics - immunology
Severe Acute Respiratory Syndrome - diagnosis - virology
Viral Envelope Proteins - genetics - immunology
 
Issue Date2005
 
PublisherAmerican Society for Microbiology.
 
CitationJournal Of Clinical Microbiology, 2005, v. 43 n. 7, p. 3054-3058 [How to Cite?]
DOI: http://dx.doi.org/10.1128/JCM.43.7.3054-3058.2005
 
AbstractThe 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. Copyright © 2005, American Society for Microbiology. All Rights Reserved.
 
ISSN0095-1137
2012 Impact Factor: 4.068
2012 SCImago Journal Rankings: 1.785
 
DOIhttp://dx.doi.org/10.1128/JCM.43.7.3054-3058.2005
 
PubMed Central IDPMC1169156
 
ISI Accession Number IDWOS:000230614900005
 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorWoo, PCY
 
dc.contributor.authorLau, SKP
 
dc.contributor.authorWong, BHL
 
dc.contributor.authorTsoi, HW
 
dc.contributor.authorFung, AMY
 
dc.contributor.authorKao, RYT
 
dc.contributor.authorChan, KH
 
dc.contributor.authorPeiris, JSM
 
dc.contributor.authorYuen, KY
 
dc.date.accessioned2008-06-12T06:35:53Z
 
dc.date.available2008-06-12T06:35:53Z
 
dc.date.issued2005
 
dc.description.abstractThe 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. Copyright © 2005, American Society for Microbiology. All Rights Reserved.
 
dc.description.naturepublished_or_final_version
 
dc.format.extent388 bytes
 
dc.format.mimetypetext/html
 
dc.identifier.citationJournal Of Clinical Microbiology, 2005, v. 43 n. 7, p. 3054-3058 [How to Cite?]
DOI: http://dx.doi.org/10.1128/JCM.43.7.3054-3058.2005
 
dc.identifier.doihttp://dx.doi.org/10.1128/JCM.43.7.3054-3058.2005
 
dc.identifier.epage3058
 
dc.identifier.hkuros109622
 
dc.identifier.isiWOS:000230614900005
 
dc.identifier.issn0095-1137
2012 Impact Factor: 4.068
2012 SCImago Journal Rankings: 1.785
 
dc.identifier.issue7
 
dc.identifier.openurl
 
dc.identifier.pmcidPMC1169156
 
dc.identifier.pmid16000415
 
dc.identifier.scopuseid_2-s2.0-22144450335
 
dc.identifier.spage3054
 
dc.identifier.urihttp://hdl.handle.net/10722/49165
 
dc.identifier.volume43
 
dc.languageeng
 
dc.publisherAmerican Society for Microbiology.
 
dc.publisher.placeUnited States
 
dc.relation.ispartofJournal of Clinical Microbiology
 
dc.relation.referencesReferences in Scopus
 
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.
 
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License
 
dc.rightsCopyright © American Society for Microbiology, Journal of Clinical Microbiology, 2005, v. 43 n. 7, p. 3054-3058
 
dc.subjectAntibodies, Viral - blood
 
dc.subjectMembrane Glycoproteins - genetics - immunology
 
dc.subjectNucleocapsid Proteins - genetics - immunology
 
dc.subjectSevere Acute Respiratory Syndrome - diagnosis - virology
 
dc.subjectViral Envelope Proteins - genetics - immunology
 
dc.titleDifferential 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
 
dc.typeArticle
 
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Author Affiliations
  1. The University of Hong Kong