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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
Lau, SKP1
Wong, BHL1
Tsoi, HW1
Fung, AMY1
Kao, RYT1
Chan, KH1
Peiris, JSM1
Yuen, KY1
 
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
2013 Impact Factor: 4.232
 
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
2013 Impact Factor: 4.232
 
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