Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA
Identifieur interne : 006002 ( Main/Merge ); précédent : 006001; suivant : 006003Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA
Auteurs : Li-Rung Huang [Taïwan] ; Chi-Ming Chiu [Taïwan] ; Shiou-Hwei Yeh [Taïwan] ; Wen-Hung Huang [Taïwan] ; Po-Ren Hsueh [Taïwan] ; Wen-Zieh Yang [Taïwan] ; Jyh-Yuan Yang [Taïwan] ; Ih-Jen Su [Taïwan] ; Shan-Chwen Chang [Taïwan] ; Pei-Jer Chenl [Taïwan]Source :
- Journal of medical virology [ 0146-6615 ] ; 2004.
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Abstract
Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. To study the humoral responses against SARS-CoV, we evaluated nucleocapsid (N) and spike (S) proteins-specific antibodies in patients' sera by Western blotting and enzyme-linked immunosorbent assay (ELISA). Recombinant N and S proteins of SARS-CoV were purified from transformed E. coli. Serum specimens from 40 SARS-CoV-infected patients in the convalescent phase were analyzed by Western blotting using the purified antigens. Serial serum specimens from 12 RT-PCR-confirmed SARS patients were assayed by ELISA using the recombinant N protein as coated antigen. By Western blotting, 97.5% of the SARS patients were positive for N protein-specific antibodies whereas only 47.5% of the samples were positive for S protein-specific antibodies. Using N protein-based ELISA, 10 out of the 12 patients were positive for N protein-specific antibodies and 6 of them showed seroconversion at mean of 16 days after onset of fever. Immunoblotting was useful for detecting the humoral immune response after SARS-CoV infection. Antibodies against SARS-CoV N protein appear at the early stage of infection, therefore, N protein-based ELISA could serve as a simple, sensitive, and specific test for diagnosing SARS-CoV infection.
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en" level="a">Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA</title><author><name sortKey="Huang, Li Rung" sort="Huang, Li Rung" uniqKey="Huang L" first="Li-Rung" last="Huang">Li-Rung Huang</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Graduate Institute of Microbiology, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Chiu, Chi Ming" sort="Chiu, Chi Ming" uniqKey="Chiu C" first="Chi-Ming" last="Chiu">Chi-Ming Chiu</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>2 aut.</sZ><sZ>4 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sort="Hsueh, Po Ren" uniqKey="Hsueh P" first="Po-Ren" last="Hsueh">Po-Ren Hsueh</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Internal Medicine, National Taiwan University Hospital</s1><s2>Taipei</s2><s3>TWN</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Wen Zieh" sort="Yang, Wen Zieh" uniqKey="Yang W" first="Wen-Zieh" last="Yang">Wen-Zieh Yang</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Center for Disease Control</s1><s2>Taipei</s2><s3>TWN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Center for Disease Control</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Jyh Yuan" sort="Yang, Jyh Yuan" uniqKey="Yang J" first="Jyh-Yuan" last="Yang">Jyh-Yuan Yang</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Center for Disease 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aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Chenl, Pei Jer" sort="Chenl, Pei Jer" uniqKey="Chenl P" first="Pei-Jer" last="Chenl">Pei-Jer Chenl</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Graduate Institute of Microbiology, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Internal Medicine, National Taiwan University Hospital</s1><s2>Taipei</s2><s3>TWN</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">04-0417134</idno><date when="2004">2004</date><idno type="stanalyst">PASCAL 04-0417134 INIST</idno><idno type="RBID">Pascal:04-0417134</idno><idno type="wicri:Area/PascalFrancis/Corpus">000834</idno><idno type="wicri:Area/PascalFrancis/Curation">000156</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000851</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000851</idno><idno type="wicri:doubleKey">0146-6615:2004:Huang L:evaluation:of:antibody</idno><idno type="wicri:Area/Main/Merge">006002</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Evaluation of antibody responses against SARS coronaviral nucleocapsid or spike proteins by immunoblotting or ELISA</title><author><name sortKey="Huang, Li Rung" sort="Huang, Li Rung" uniqKey="Huang L" first="Li-Rung" last="Huang">Li-Rung Huang</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Graduate Institute of Microbiology, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Chiu, Chi Ming" sort="Chiu, Chi Ming" uniqKey="Chiu C" first="Chi-Ming" last="Chiu">Chi-Ming Chiu</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Yeh, Shiou Hwei" sort="Yeh, Shiou Hwei" uniqKey="Yeh S" first="Shiou-Hwei" last="Yeh">Shiou-Hwei Yeh</name><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Division of Molecular and Genomic Medicine, National Health Research Institutes</s1><s2>Taipei</s2><s3>TWN</s3><sZ>3 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Huang, Wen Hung" sort="Huang, Wen Hung" uniqKey="Huang W" first="Wen-Hung" last="Huang">Wen-Hung Huang</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>2 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Hsueh, Po Ren" sort="Hsueh, Po Ren" uniqKey="Hsueh P" first="Po-Ren" last="Hsueh">Po-Ren Hsueh</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Internal Medicine, National Taiwan University Hospital</s1><s2>Taipei</s2><s3>TWN</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Wen Zieh" sort="Yang, Wen Zieh" uniqKey="Yang W" first="Wen-Zieh" last="Yang">Wen-Zieh Yang</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Center for Disease Control</s1><s2>Taipei</s2><s3>TWN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Center for Disease Control</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Jyh Yuan" sort="Yang, Jyh Yuan" uniqKey="Yang J" first="Jyh-Yuan" last="Yang">Jyh-Yuan Yang</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Center for Disease Control</s1><s2>Taipei</s2><s3>TWN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Center for Disease Control</wicri:noRegion></affiliation></author><author><name sortKey="Su, Ih Jen" sort="Su, Ih Jen" uniqKey="Su I" first="Ih-Jen" last="Su">Ih-Jen Su</name><affiliation wicri:level="1"><inist:fA14 i1="05"><s1>Center for Disease Control</s1><s2>Taipei</s2><s3>TWN</s3><sZ>6 aut.</sZ><sZ>7 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Center for Disease Control</wicri:noRegion></affiliation></author><author><name sortKey="Chang, Shan Chwen" sort="Chang, Shan Chwen" uniqKey="Chang S" first="Shan-Chwen" last="Chang">Shan-Chwen Chang</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Internal Medicine, National Taiwan University Hospital</s1><s2>Taipei</s2><s3>TWN</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author><author><name sortKey="Chenl, Pei Jer" sort="Chenl, Pei Jer" uniqKey="Chenl P" first="Pei-Jer" last="Chenl">Pei-Jer Chenl</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Graduate Institute of Microbiology, National Taiwan University College of Medicine</s1><s2>Taipei</s2><s3>TWN</s3><sZ>1 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Internal Medicine, National Taiwan University Hospital</s1><s2>Taipei</s2><s3>TWN</s3><sZ>5 aut.</sZ><sZ>9 aut.</sZ><sZ>10 aut.</sZ></inist:fA14><country>Taïwan</country><wicri:noRegion>Taipei</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of medical virology</title><title level="j" type="abbreviated">J. med. virol.</title><idno type="ISSN">0146-6615</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Antibody</term><term>ELISA assay</term><term>Immunoblotting assay</term><term>Nucleocapsid</term><term>Protein</term><term>Severe acute respiratory syndrome</term><term>Vitronectin</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Anticorps</term><term>Nucléocapside</term><term>Protéine</term><term>Méthode immunoblotting</term><term>Technique ELISA</term><term>Vitronectine</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome (SARS)-CoV is a newly emerging virus that causes SARS with high mortality rate in infected people. To study the humoral responses against SARS-CoV, we evaluated nucleocapsid (N) and spike (S) proteins-specific antibodies in patients' sera by Western blotting and enzyme-linked immunosorbent assay (ELISA). Recombinant N and S proteins of SARS-CoV were purified from transformed E. coli. Serum specimens from 40 SARS-CoV-infected patients in the convalescent phase were analyzed by Western blotting using the purified antigens. Serial serum specimens from 12 RT-PCR-confirmed SARS patients were assayed by ELISA using the recombinant N protein as coated antigen. By Western blotting, 97.5% of the SARS patients were positive for N protein-specific antibodies whereas only 47.5% of the samples were positive for S protein-specific antibodies. Using N protein-based ELISA, 10 out of the 12 patients were positive for N protein-specific antibodies and 6 of them showed seroconversion at mean of 16 days after onset of fever. Immunoblotting was useful for detecting the humoral immune response after SARS-CoV infection. Antibodies against SARS-CoV N protein appear at the early stage of infection, therefore, N protein-based ELISA could serve as a simple, sensitive, and specific test for diagnosing SARS-CoV infection.</div></front></TEI><affiliations><list><country><li>Taïwan</li></country></list><tree><country name="Taïwan"><noRegion><name sortKey="Huang, Li Rung" sort="Huang, Li Rung" uniqKey="Huang L" first="Li-Rung" last="Huang">Li-Rung Huang</name></noRegion><name sortKey="Chang, Shan Chwen" sort="Chang, Shan Chwen" uniqKey="Chang S" first="Shan-Chwen" last="Chang">Shan-Chwen Chang</name><name sortKey="Chenl, Pei Jer" sort="Chenl, Pei Jer" uniqKey="Chenl P" first="Pei-Jer" last="Chenl">Pei-Jer Chenl</name><name sortKey="Chenl, Pei Jer" sort="Chenl, Pei Jer" uniqKey="Chenl P" first="Pei-Jer" last="Chenl">Pei-Jer Chenl</name><name sortKey="Chiu, Chi Ming" sort="Chiu, Chi Ming" uniqKey="Chiu C" first="Chi-Ming" last="Chiu">Chi-Ming Chiu</name><name sortKey="Hsueh, Po Ren" sort="Hsueh, Po Ren" uniqKey="Hsueh P" first="Po-Ren" last="Hsueh">Po-Ren Hsueh</name><name sortKey="Huang, Wen Hung" sort="Huang, Wen Hung" uniqKey="Huang W" first="Wen-Hung" last="Huang">Wen-Hung Huang</name><name sortKey="Su, Ih Jen" sort="Su, Ih Jen" uniqKey="Su I" first="Ih-Jen" last="Su">Ih-Jen Su</name><name sortKey="Yang, Jyh Yuan" sort="Yang, Jyh Yuan" uniqKey="Yang J" first="Jyh-Yuan" last="Yang">Jyh-Yuan Yang</name><name sortKey="Yang, Wen Zieh" sort="Yang, Wen Zieh" uniqKey="Yang W" first="Wen-Zieh" last="Yang">Wen-Zieh Yang</name><name sortKey="Yeh, Shiou Hwei" sort="Yeh, Shiou Hwei" uniqKey="Yeh S" first="Shiou-Hwei" last="Yeh">Shiou-Hwei Yeh</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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