A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses
Identifieur interne : 000F24 ( Pmc/Curation ); précédent : 000F23; suivant : 000F25A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses
Auteurs : Christian Lehmann [Allemagne] ; Hans Wolf [Allemagne] ; Jianguo Xu [République populaire de Chine] ; Quanbi Zhao [République populaire de Chine] ; Yiming Shao [République populaire de Chine] ; Manfred Motz [Allemagne] ; Petra Lindner [Allemagne]Source :
- Diagnostic Microbiology and Infectious Disease [ 0732-8893 ] ; 2008.
Abstract
Most coronaviruses infecting humans cause mild diseases, whereas severe acute respiratory syndrome (SARS)-associated coronavirus is an extremely dangerous pathogen. Here, we report the development of a serologic assay for detection of antibodies to human coronaviruses (HCoVs) based on recombinant nucleocapsid (N) proteins of all known pathogenic strains (229E, NL63, OC43, HKU1, SARS). The novel immunoassay is highly useful for epidemiologic surveys, where use of nucleic acid diagnostics often is limited. Purified recombinant antigens were immobilized on nitrocellulose membranes and applied in a line immunoassay, which allows rapid detection of antibodies to 5 different HCoVs in a single experiment. For assay evaluation, serum samples from persons infected with 229E or OC43 (acute/convalescent), recovered SARS patients and healthy donors were analyzed. Screening for nucleocapsid (N)-specific immunoglobulin G (IgG) in convalescent sera reached 100% sensitivity. With this new technique, we found that recently identified NL63 and HKU1 contribute significantly to the overall spectrum of coronavirus infections. Possibly, cross-reactive antibody responses were observed using 229E and OC43 serum pairs. However, the potential of this assay could clearly be demonstrated employing SARS-positive serum samples, where nonspecific binding to nucleocapsids of other HCoVs was not observed. This coronavirus strain-specific line immunoassay represents a powerful tool for serologic diagnostics.
Url:
DOI: 10.1016/j.diagmicrobio.2007.12.002
PubMed: 18191362
PubMed Central: 7127592
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wicri:step="Curation">000F24</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses</title><author><name sortKey="Lehmann, Christian" sort="Lehmann, Christian" uniqKey="Lehmann C" first="Christian" last="Lehmann">Christian Lehmann</name><affiliation wicri:level="1"><nlm:aff id="aff1">Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg</wicri:regionArea></affiliation></author><author><name sortKey="Wolf, Hans" sort="Wolf, Hans" uniqKey="Wolf H" first="Hans" last="Wolf">Hans Wolf</name><affiliation wicri:level="1"><nlm:aff id="aff1">Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg</wicri:regionArea></affiliation></author><author><name sortKey="Xu, Jianguo" sort="Xu, Jianguo" uniqKey="Xu J" first="Jianguo" last="Xu">Jianguo Xu</name><affiliation wicri:level="1"><nlm:aff id="aff2">Chinese Center for Disease Control and Prevention (China CDC), Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Center for Disease Control and Prevention (China CDC), Beijing</wicri:regionArea></affiliation></author><author><name sortKey="Zhao, Quanbi" sort="Zhao, Quanbi" uniqKey="Zhao Q" first="Quanbi" last="Zhao">Quanbi Zhao</name><affiliation wicri:level="1"><nlm:aff id="aff2">Chinese Center for Disease Control and Prevention (China CDC), Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Center for Disease Control and Prevention (China CDC), Beijing</wicri:regionArea></affiliation></author><author><name sortKey="Shao, Yiming" sort="Shao, Yiming" uniqKey="Shao Y" first="Yiming" last="Shao">Yiming Shao</name><affiliation wicri:level="1"><nlm:aff id="aff2">Chinese Center for Disease Control and Prevention (China CDC), Beijing, PR China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Center for Disease Control and Prevention (China CDC), Beijing</wicri:regionArea></affiliation></author><author><name sortKey="Motz, Manfred" sort="Motz, Manfred" uniqKey="Motz M" first="Manfred" last="Motz">Manfred Motz</name><affiliation wicri:level="1"><nlm:aff id="aff3">Mikrogen GmbH, D-82061 Neuried, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Mikrogen GmbH, D-82061 Neuried</wicri:regionArea></affiliation></author><author><name sortKey="Lindner, Petra" sort="Lindner, Petra" uniqKey="Lindner P" first="Petra" last="Lindner">Petra Lindner</name><affiliation wicri:level="1"><nlm:aff id="aff1">Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany</nlm:aff><country xml:lang="fr">Allemagne</country><wicri:regionArea>Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg</wicri:regionArea></affiliation></author></analytic><series><title level="j">Diagnostic Microbiology and Infectious Disease</title><idno type="ISSN">0732-8893</idno><idno type="eISSN">1879-0070</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Most coronaviruses infecting humans cause mild diseases, whereas severe acute respiratory syndrome (SARS)-associated coronavirus is an extremely dangerous pathogen. Here, we report the development of a serologic assay for detection of antibodies to human coronaviruses (HCoVs) based on recombinant nucleocapsid (N) proteins of all known pathogenic strains (229E, NL63, OC43, HKU1, SARS). The novel immunoassay is highly useful for epidemiologic surveys, where use of nucleic acid diagnostics often is limited. Purified recombinant antigens were immobilized on nitrocellulose membranes and applied in a line immunoassay, which allows rapid detection of antibodies to 5 different HCoVs in a single experiment. For assay evaluation, serum samples from persons infected with 229E or OC43 (acute/convalescent), recovered SARS patients and healthy donors were analyzed. Screening for nucleocapsid (N)-specific immunoglobulin G (IgG) in convalescent sera reached 100% sensitivity. With this new technique, we found that recently identified NL63 and HKU1 contribute significantly to the overall spectrum of coronavirus infections. Possibly, cross-reactive antibody responses were observed using 229E and OC43 serum pairs. However, the potential of this assay could clearly be demonstrated employing SARS-positive serum samples, where nonspecific binding to nucleocapsids of other HCoVs was not observed. This coronavirus strain-specific line immunoassay represents a powerful tool for serologic diagnostics.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Arden, K E" uniqKey="Arden K">K.E. Arden</name></author><author><name sortKey="Nissen, M D" uniqKey="Nissen M">M.D. Nissen</name></author><author><name sortKey="Sloots, T P" uniqKey="Sloots T">T.P. Sloots</name></author><author><name sortKey="Sloots, T P" uniqKey="Sloots T">T.P. Sloots</name></author><author><name sortKey="Mackay, I M" uniqKey="Mackay I">I.M. Mackay</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bastien, N" uniqKey="Bastien N">N. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Diagn Microbiol Infect Dis</journal-id><journal-id journal-id-type="iso-abbrev">Diagn. Microbiol. Infect. Dis</journal-id><journal-title-group><journal-title>Diagnostic Microbiology and Infectious Disease</journal-title></journal-title-group><issn pub-type="ppub">0732-8893</issn><issn pub-type="epub">1879-0070</issn><publisher><publisher-name>Elsevier Inc.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">18191362</article-id><article-id pub-id-type="pmc">7127592</article-id><article-id pub-id-type="publisher-id">S0732-8893(07)00588-3</article-id><article-id pub-id-type="doi">10.1016/j.diagmicrobio.2007.12.002</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>A line immunoassay utilizing recombinant nucleocapsid proteins for detection of antibodies to human coronaviruses</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Lehmann</surname><given-names>Christian</given-names></name><email>christian.lehmann@klinik.uni-r.de</email><xref rid="aff1" ref-type="aff">a</xref><xref rid="cor1" ref-type="corresp">⁎</xref></contrib><contrib contrib-type="author"><name><surname>Wolf</surname><given-names>Hans</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib><contrib contrib-type="author"><name><surname>Xu</surname><given-names>Jianguo</given-names></name><xref rid="aff2" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Zhao</surname><given-names>Quanbi</given-names></name><xref rid="aff2" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Shao</surname><given-names>Yiming</given-names></name><xref rid="aff2" ref-type="aff">b</xref></contrib><contrib contrib-type="author"><name><surname>Motz</surname><given-names>Manfred</given-names></name><xref rid="aff3" ref-type="aff">c</xref></contrib><contrib contrib-type="author"><name><surname>Lindner</surname><given-names>Petra</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib></contrib-group><aff id="aff1"><label>a</label>Institute for Medical Microbiology and Hygiene, University of Regensburg, 93053 Regensburg, Germany</aff><aff id="aff2"><label>b</label>Chinese Center for Disease Control and Prevention (China CDC), Beijing, PR China</aff><aff id="aff3"><label>c</label>Mikrogen GmbH, D-82061 Neuried, Germany</aff><author-notes><corresp id="cor1"><label>⁎</label>Corresponding author. Tel.: +49-941-944-4628; fax: +49-941-944-6402. <email>christian.lehmann@klinik.uni-r.de</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>11</day><month>1</month><year>2008</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>5</month><year>2008</year></pub-date><pub-date pub-type="epub"><day>11</day><month>1</month><year>2008</year></pub-date><volume>61</volume><issue>1</issue><fpage>40</fpage><lpage>48</lpage><history><date date-type="received"><day>20</day><month>6</month><year>2007</year></date><date date-type="accepted"><day>3</day><month>12</month><year>2007</year></date></history><permissions><copyright-statement>Copyright © 2008 Elsevier Inc. All rights reserved.</copyright-statement><copyright-year>2008</copyright-year><copyright-holder>Elsevier Inc.</copyright-holder><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract><p>Most coronaviruses infecting humans cause mild diseases, whereas severe acute respiratory syndrome (SARS)-associated coronavirus is an extremely dangerous pathogen. Here, we report the development of a serologic assay for detection of antibodies to human coronaviruses (HCoVs) based on recombinant nucleocapsid (N) proteins of all known pathogenic strains (229E, NL63, OC43, HKU1, SARS). The novel immunoassay is highly useful for epidemiologic surveys, where use of nucleic acid diagnostics often is limited. Purified recombinant antigens were immobilized on nitrocellulose membranes and applied in a line immunoassay, which allows rapid detection of antibodies to 5 different HCoVs in a single experiment. For assay evaluation, serum samples from persons infected with 229E or OC43 (acute/convalescent), recovered SARS patients and healthy donors were analyzed. Screening for nucleocapsid (N)-specific immunoglobulin G (IgG) in convalescent sera reached 100% sensitivity. With this new technique, we found that recently identified NL63 and HKU1 contribute significantly to the overall spectrum of coronavirus infections. Possibly, cross-reactive antibody responses were observed using 229E and OC43 serum pairs. However, the potential of this assay could clearly be demonstrated employing SARS-positive serum samples, where nonspecific binding to nucleocapsids of other HCoVs was not observed. This coronavirus strain-specific line immunoassay represents a powerful tool for serologic diagnostics.</p></abstract><kwd-group><title>Keywords</title><kwd>Human coronavirus</kwd><kwd>Nucleocapsid</kwd><kwd>Line immunoassay</kwd><kwd>SARS</kwd><kwd>Seroprevalence</kwd><kwd>Cross-reaction</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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