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
2011 Impact Factor: 4.153
2011 SCImago Journal Rankings: 0.397
DOIhttp://dx.doi.org/10.1128/JCM.43.7.3054-3058.2005
ISI Accession Number IDWOS:000230614900005
PubMed Central IDPMC1169156
ReferencesReferences in Scopus
DC Field
Value
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
2011 Impact Factor: 4.153
2011 SCImago Journal Rankings: 0.397
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
Author Affiliations
  1. The University of Hong Kong