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Article: Sensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndrome

TitleSensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndrome
Authors
Issue Date2004
PublisherAmerican Society for Microbiology.
Citation
Journal Of Clinical Microbiology, 2004, v. 42 n. 6, p. 2629-2635 How to Cite?
AbstractA rapid antigen test for the diagnosis of severe acute respiratory syndrome (SARS) is essential for control of this disease at the point of management. The nucleocapsid (N) protein of SARS-associated coronavirus (SARS-CoV) is abundantly expressed in infected-cell culture filtrate as demonstrable by Western blotting using convalescent-phase sera from patients with SARS. We used monoclonal antibodies specifically directed against N protein to establish a sensitive antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of SARS-CoV. The assay employed a mixture of three monoclonal antibodies for capture and rabbit polyclonal antibodies for detection of serum antigen in 32 cases of clinically probable SARS as defined by the World Health Organization during the epidemic in Guangzhou, China. Recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml. The linear range of detection in clinical specimens was from 100 pg/ml to 3.2 ng/ml. Using a panel of sera collected at different points in time, the amount of circulating N antigen was found to peak 6 to 10 days after the onset of symptoms. The sensitivity of the assay was 84.6% in 13 serologically confirmed SARS patients with blood taken during the first 10 days after the onset of symptoms (11 of 13). The specificity of the assay was 98.5% in 1,272 healthy individuals (1,253 of 1,272). There was no cross-reaction with other human and animal coronaviruses in this assay. In conclusion, a sensitive and quantitative antigen capture ELISA was established for the early diagnosis and disease monitoring of SARS-CoV infection.
Persistent Identifierhttp://hdl.handle.net/10722/49259
ISSN
2015 Impact Factor: 3.631
2015 SCImago Journal Rankings: 2.151
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorChe, XYen_HK
dc.contributor.authorQiu, LWen_HK
dc.contributor.authorPan, YXen_HK
dc.contributor.authorWen, Ken_HK
dc.contributor.authorHao, Wen_HK
dc.contributor.authorZhang, LYen_HK
dc.contributor.authorWang, YDen_HK
dc.contributor.authorLiao, ZYen_HK
dc.contributor.authorHua, Xen_HK
dc.contributor.authorCheng, VCCen_HK
dc.contributor.authorYuen, KYen_HK
dc.date.accessioned2008-06-12T06:37:46Z-
dc.date.available2008-06-12T06:37:46Z-
dc.date.issued2004en_HK
dc.identifier.citationJournal Of Clinical Microbiology, 2004, v. 42 n. 6, p. 2629-2635en_HK
dc.identifier.issn0095-1137en_HK
dc.identifier.urihttp://hdl.handle.net/10722/49259-
dc.description.abstractA rapid antigen test for the diagnosis of severe acute respiratory syndrome (SARS) is essential for control of this disease at the point of management. The nucleocapsid (N) protein of SARS-associated coronavirus (SARS-CoV) is abundantly expressed in infected-cell culture filtrate as demonstrable by Western blotting using convalescent-phase sera from patients with SARS. We used monoclonal antibodies specifically directed against N protein to establish a sensitive antigen capture sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of SARS-CoV. The assay employed a mixture of three monoclonal antibodies for capture and rabbit polyclonal antibodies for detection of serum antigen in 32 cases of clinically probable SARS as defined by the World Health Organization during the epidemic in Guangzhou, China. Recombinant N protein was used as a standard to establish a detection sensitivity of approximated 50 pg/ml. The linear range of detection in clinical specimens was from 100 pg/ml to 3.2 ng/ml. Using a panel of sera collected at different points in time, the amount of circulating N antigen was found to peak 6 to 10 days after the onset of symptoms. The sensitivity of the assay was 84.6% in 13 serologically confirmed SARS patients with blood taken during the first 10 days after the onset of symptoms (11 of 13). The specificity of the assay was 98.5% in 1,272 healthy individuals (1,253 of 1,272). There was no cross-reaction with other human and animal coronaviruses in this assay. In conclusion, a sensitive and quantitative antigen capture ELISA was established for the early diagnosis and disease monitoring of SARS-CoV infection.en_HK
dc.format.extent386 bytes-
dc.format.mimetypetext/html-
dc.languageengen_HK
dc.publisherAmerican Society for Microbiology.en_HK
dc.relation.ispartofJournal of Clinical Microbiologyen_HK
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.rightsJournal of Clinical Microbiology. Copyright © American Society for Microbiology.en_HK
dc.rightsCopyright © American Society for Microbiology, Journal of Clinical Microbiology, 2004, v. 42 n. 6, p. 2629-2635en_HK
dc.subject.meshAntibodies, Monoclonal - immunologyen_HK
dc.subject.meshAntigens, Viral - blooden_HK
dc.subject.meshNucleocapsid - blooden_HK
dc.subject.meshSARS Virus - immunologyen_HK
dc.subject.meshSevere Acute Respiratory Syndrome - diagnosisen_HK
dc.titleSensitive and specific monoclonal antibody-based capture enzyme immunoassay for detection of nucleocapsid antigen in sera from patients with severe acute respiratory syndromeen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0095-1137&volume=42&issue=6&spage=2629&epage=2635&date=2004&atitle=Sensitive+and+specific+monoclonal+antibody-based+capture+enzyme+immunoassay+for+detection+of+nucleocapsid+antigen+in+sera+from+patients+with+severe+acute+respiratory+syndrome.+en_HK
dc.identifier.emailYuen, KY:kyyuen@hkucc.hku.hken_HK
dc.identifier.authorityYuen, KY=rp00366en_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.1128/JCM.42.6.2629-2635.2004en_HK
dc.identifier.pmid15184444-
dc.identifier.pmcidPMC427886en_HK
dc.identifier.scopuseid_2-s2.0-2942568032en_HK
dc.identifier.hkuros95885-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-2942568032&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume42en_HK
dc.identifier.issue6en_HK
dc.identifier.spage2629en_HK
dc.identifier.epage2635en_HK
dc.identifier.isiWOS:000222015800040-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridChe, XY=7005743182en_HK
dc.identifier.scopusauthoridQiu, LW=8719102700en_HK
dc.identifier.scopusauthoridPan, YX=35765899800en_HK
dc.identifier.scopusauthoridWen, K=8231190100en_HK
dc.identifier.scopusauthoridHao, W=7101686587en_HK
dc.identifier.scopusauthoridZhang, LY=37086714000en_HK
dc.identifier.scopusauthoridWang, YD=9638471800en_HK
dc.identifier.scopusauthoridLiao, ZY=7203032864en_HK
dc.identifier.scopusauthoridHua, X=36817185600en_HK
dc.identifier.scopusauthoridCheng, VCC=23670479400en_HK
dc.identifier.scopusauthoridYuen, KY=36078079100en_HK

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