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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Development of a Western Blot Assay for Detection of Antibodies against Coronavirus Causing Severe Acute Respiratory Syndrome</title><author><name sortKey="He, Qigai" sort="He, Qigai" uniqKey="He Q" first="Qigai" last="He">Qigai He</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chong, Kooi Hoong" sort="Chong, Kooi Hoong" uniqKey="Chong K" first="Kooi Hoong" last="Chong">Kooi Hoong Chong</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hee Chng, Hiok" sort="Hee Chng, Hiok" uniqKey="Hee Chng H" first="Hiok" last="Hee Chng">Hiok Hee Chng</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Leung, Bernard" sort="Leung, Bernard" uniqKey="Leung B" first="Bernard" last="Leung">Bernard Leung</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ee Ling, Ai" sort="Ee Ling, Ai" uniqKey="Ee Ling A" first="Ai" last="Ee Ling">Ai Ee Ling</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Wei, Ting" sort="Wei, Ting" uniqKey="Wei T" first="Ting" last="Wei">Ting Wei</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Eong Ooi, Eng" sort="Eong Ooi, Eng" uniqKey="Eong Ooi E" first="Eng" last="Eong Ooi">Eng Eong Ooi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15013997</idno><idno type="pmc">371214</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC371214</idno><idno type="RBID">PMC:371214</idno><idno type="doi">10.1128/CDLI.11.2.417-422.2004</idno><date when="2004">2004</date><idno type="wicri:Area/Pmc/Corpus">000627</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000627</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Development of a Western Blot Assay for Detection of Antibodies against Coronavirus Causing Severe Acute Respiratory Syndrome</title><author><name sortKey="He, Qigai" sort="He, Qigai" uniqKey="He Q" first="Qigai" last="He">Qigai He</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chong, Kooi Hoong" sort="Chong, Kooi Hoong" uniqKey="Chong K" first="Kooi Hoong" last="Chong">Kooi Hoong Chong</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hee Chng, Hiok" sort="Hee Chng, Hiok" uniqKey="Hee Chng H" first="Hiok" last="Hee Chng">Hiok Hee Chng</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Leung, Bernard" sort="Leung, Bernard" uniqKey="Leung B" first="Bernard" last="Leung">Bernard Leung</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ee Ling, Ai" sort="Ee Ling, Ai" uniqKey="Ee Ling A" first="Ai" last="Ee Ling">Ai Ee Ling</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Wei, Ting" sort="Wei, Ting" uniqKey="Wei T" first="Ting" last="Wei">Ting Wei</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Chan, Shzu Wei" sort="Chan, Shzu Wei" uniqKey="Chan S" first="Shzu-Wei" last="Chan">Shzu-Wei Chan</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Eong Ooi, Eng" sort="Eong Ooi, Eng" uniqKey="Eong Ooi E" first="Eng" last="Eong Ooi">Eng Eong Ooi</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kwang, Jimmy" sort="Kwang, Jimmy" uniqKey="Kwang J" first="Jimmy" last="Kwang">Jimmy Kwang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Clinical and Diagnostic Laboratory Immunology</title><idno type="ISSN">1071-412X</idno><idno type="eISSN">1098-6588</idno><imprint><date when="2004">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>To identify a major antigenic determinant for use in the development of a rapid serological diagnostic test for severe acute respiratory syndrome (SARS) coronavirus infection and to study the immune response during SARS coronavirus infection in humans, we cloned the full length and six truncated fragments of the nucleocapsid gene, expressed them, and purified them as glutathione <italic>S</italic>-transferase-tagged recombinant proteins. The reactivities of the recombinant proteins to a panel of antibodies containing 33 SARS coronavirus-positive sera and 66 negative sera and to antibodies against other animal coronaviruses were screened. A truncated 195-amino-acid fragment from the C terminus of the nucleocapsid protein (N195) was identified that had a strong ability to detect antibodies against SARS coronavirus. No cross-reaction was found between the N195 protein and antibodies against chicken, pig, and canine coronaviruses. The N195 protein was used to develop a Western blot assay to detect antibodies against SARS coronavirus in 274 clinically blinded samples. The specificity and sensitivity of this test were 98.3 and 90.9%, respectively. The correlation between our Western blotting assay and an immunofluorescence assay (IFA) was also analyzed. The results of our Western blot assay and IFA for the detection of SARS coronavirus-positive sera were the same. Thus, the N195 protein was identified as a suitable protein to be used as an antigen in Western blot and other possible assays for the detection of SARS coronavirus infection.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Clin Diagn Lab Immunol</journal-id><journal-id journal-id-type="publisher-id">cdli</journal-id><journal-title>Clinical and Diagnostic Laboratory Immunology</journal-title><issn pub-type="ppub">1071-412X</issn><issn pub-type="epub">1098-6588</issn><publisher><publisher-name>American Society for Microbiology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15013997</article-id><article-id pub-id-type="pmc">371214</article-id><article-id pub-id-type="publisher-id">0226</article-id><article-id pub-id-type="doi">10.1128/CDLI.11.2.417-422.2004</article-id><article-categories><subj-group subj-group-type="heading"><subject>Microbial Immunology</subject></subj-group></article-categories><title-group><article-title>Development of a Western Blot Assay for Detection of Antibodies against Coronavirus Causing Severe Acute Respiratory Syndrome</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>He</surname><given-names>Qigai</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Chong</surname><given-names>Kooi Hoong</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Hee Chng</surname><given-names>Hiok</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Leung</surname><given-names>Bernard</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Ee Ling</surname><given-names>Ai</given-names></name><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Wei</surname><given-names>Ting</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Chan</surname><given-names>Shzu-Wei</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Eong Ooi</surname><given-names>Eng</given-names></name><xref ref-type="aff" rid="aff1">4</xref></contrib><contrib contrib-type="author"><name><surname>Kwang</surname><given-names>Jimmy</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Animal Health Biotechnology, Temasek Life Science Laboratory, National University of Singapore,<label>1</label> Department of Rheumatology, Allergy and Immunology, Tan Tock Seng Hospital,<label>2</label> Virology Section, Department of Pathology, Singapore General Hospital,<label>3</label> Environment Health Institute, National Environment Agency, Singapore, Republic of Singapore<label>4</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Animal Health Biotechnology, Temasek Life Science Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Republic of Singapore. Phone: 65-68727473. Fax: 65-68727007. E-mail: <email>kwang@tll.org.sg</email>.</p></fn></author-notes><pub-date pub-type="ppub"><month>3</month><year>2004</year></pub-date><volume>11</volume><issue>2</issue><fpage>417</fpage><lpage>422</lpage><history><date date-type="received"><day>23</day><month>9</month><year>2003</year></date><date date-type="rev-recd"><day>4</day><month>11</month><year>2003</year></date><date date-type="accepted"><day>4</day><month>12</month><year>2003</year></date></history><copyright-statement>Copyright © 2004, American Society for Microbiology</copyright-statement><copyright-year>2004</copyright-year><abstract><p>To identify a major antigenic determinant for use in the development of a rapid serological diagnostic test for severe acute respiratory syndrome (SARS) coronavirus infection and to study the immune response during SARS coronavirus infection in humans, we cloned the full length and six truncated fragments of the nucleocapsid gene, expressed them, and purified them as glutathione <italic>S</italic>-transferase-tagged recombinant proteins. The reactivities of the recombinant proteins to a panel of antibodies containing 33 SARS coronavirus-positive sera and 66 negative sera and to antibodies against other animal coronaviruses were screened. A truncated 195-amino-acid fragment from the C terminus of the nucleocapsid protein (N195) was identified that had a strong ability to detect antibodies against SARS coronavirus. No cross-reaction was found between the N195 protein and antibodies against chicken, pig, and canine coronaviruses. The N195 protein was used to develop a Western blot assay to detect antibodies against SARS coronavirus in 274 clinically blinded samples. The specificity and sensitivity of this test were 98.3 and 90.9%, respectively. The correlation between our Western blotting assay and an immunofluorescence assay (IFA) was also analyzed. The results of our Western blot assay and IFA for the detection of SARS coronavirus-positive sera were the same. Thus, the N195 protein was identified as a suitable protein to be used as an antigen in Western blot and other possible assays for the detection of SARS coronavirus infection.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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