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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

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
Authors
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.
Citation
Journal Of Clinical Microbiology, 2005, v. 43 n. 7, p. 3054-3058 How to Cite?
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.
Persistent Identifierhttp://hdl.handle.net/10722/49165
ISSN
2014 Impact Factor: 3.993
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorWoo, PCYen_HK
dc.contributor.authorLau, SKPen_HK
dc.contributor.authorWong, BHLen_HK
dc.contributor.authorTsoi, HWen_HK
dc.contributor.authorFung, AMYen_HK
dc.contributor.authorKao, RYTen_HK
dc.contributor.authorChan, KHen_HK
dc.contributor.authorPeiris, JSMen_HK
dc.contributor.authorYuen, KYen_HK
dc.date.accessioned2008-06-12T06:35:53Z-
dc.date.available2008-06-12T06:35:53Z-
dc.date.issued2005en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2005, v. 43 n. 7, p. 3054-3058en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49165-
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.en_HK
dc.format.extent388 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.en_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
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. 3054-3058en_HK
dc.subjectAntibodies, Viral - blooden_HK
dc.subjectMembrane Glycoproteins - genetics - immunologyen_HK
dc.subjectNucleocapsid Proteins - genetics - immunologyen_HK
dc.subjectSevere Acute Respiratory Syndrome - diagnosis - virologyen_HK
dc.subjectViral Envelope Proteins - genetics - immunologyen_HK
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 pneumoniaen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=43&issue=7&spage=3054&epage=3058&date=2005&atitle=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+pneumoniaen_HK
dc.identifier.emailWoo, PCY:pcywoo@hkucc.hku.hken_HK
dc.identifier.emailLau, SKP:skplau@hkucc.hku.hken_HK
dc.identifier.emailTsoi, HW:hwtsoi@hkucc.hku.hken_HK
dc.identifier.emailKao, RYT:rytkao@hkucc.hku.hken_HK
dc.identifier.emailPeiris, JSM:malik@hkucc.hku.hken_HK
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_HK
dc.identifier.authorityWoo, PCY=rp00430en_HK
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dc.identifier.authorityTsoi, HW=rp00439en_HK
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dc.identifier.authorityPeiris, JSM=rp00410en_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1128/JCM.43.7.3054-3058.2005en_HK
dc.identifier.pmid16000415en_HK
dc.identifier.pmcidPMC1169156en_HK
dc.identifier.scopuseid_2-s2.0-22144450335en_HK
dc.identifier.hkuros109622-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-22144450335&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume43en_HK
dc.identifier.issue7en_HK
dc.identifier.spage3054en_HK
dc.identifier.epage3058en_HK
dc.identifier.isiWOS:000230614900005-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridWoo, PCY=7201801340en_HK
dc.identifier.scopusauthoridLau, SKP=7401596211en_HK
dc.identifier.scopusauthoridWong, BHL=7402023413en_HK
dc.identifier.scopusauthoridTsoi, HW=6603822102en_HK
dc.identifier.scopusauthoridFung, AMY=7101926801en_HK
dc.identifier.scopusauthoridKao, RYT=7101675499en_HK
dc.identifier.scopusauthoridChan, KH=7406034307en_HK
dc.identifier.scopusauthoridPeiris, JSM=7005486823en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK

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