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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Two-Way Antigenic Cross-Reactivity between Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Group 1 Animal CoVs Is Mediated through an Antigenic Site in the N-Terminal Region of the SARS-CoV Nucleoprotein<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Vlasova, Anastasia N" sort="Vlasova, Anastasia N" uniqKey="Vlasova A" first="Anastasia N." last="Vlasova">Anastasia N. Vlasova</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Zhang, Xinsheng" sort="Zhang, Xinsheng" uniqKey="Zhang X" first="Xinsheng" last="Zhang">Xinsheng Zhang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hasoksuz, Mustafa" sort="Hasoksuz, Mustafa" uniqKey="Hasoksuz M" first="Mustafa" last="Hasoksuz">Mustafa Hasoksuz</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Nagesha, Hadya S" sort="Nagesha, Hadya S" uniqKey="Nagesha H" first="Hadya S." last="Nagesha">Hadya S. Nagesha</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Haynes, Lia M" sort="Haynes, Lia M" uniqKey="Haynes L" first="Lia M." last="Haynes">Lia M. Haynes</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Fang, Ying" sort="Fang, Ying" uniqKey="Fang Y" first="Ying" last="Fang">Ying Fang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lu, Shan" sort="Lu, Shan" uniqKey="Lu S" first="Shan" last="Lu">Shan Lu</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Saif, Linda J" sort="Saif, Linda J" uniqKey="Saif L" first="Linda J." last="Saif">Linda J. Saif</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">17913799</idno><idno type="pmc">2168854</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2168854</idno><idno type="RBID">PMC:2168854</idno><idno type="doi">10.1128/JVI.01169-07</idno><date when="2007">2007</date><idno type="wicri:Area/Pmc/Corpus">000723</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000723</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Two-Way Antigenic Cross-Reactivity between Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Group 1 Animal CoVs Is Mediated through an Antigenic Site in the N-Terminal Region of the SARS-CoV Nucleoprotein<xref ref-type="fn" rid="fn1">▿</xref> </title><author><name sortKey="Vlasova, Anastasia N" sort="Vlasova, Anastasia N" uniqKey="Vlasova A" first="Anastasia N." last="Vlasova">Anastasia N. Vlasova</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Zhang, Xinsheng" sort="Zhang, Xinsheng" uniqKey="Zhang X" first="Xinsheng" last="Zhang">Xinsheng Zhang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Hasoksuz, Mustafa" sort="Hasoksuz, Mustafa" uniqKey="Hasoksuz M" first="Mustafa" last="Hasoksuz">Mustafa Hasoksuz</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Nagesha, Hadya S" sort="Nagesha, Hadya S" uniqKey="Nagesha H" first="Hadya S." last="Nagesha">Hadya S. Nagesha</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Haynes, Lia M" sort="Haynes, Lia M" uniqKey="Haynes L" first="Lia M." last="Haynes">Lia M. Haynes</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Fang, Ying" sort="Fang, Ying" uniqKey="Fang Y" first="Ying" last="Fang">Ying Fang</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lu, Shan" sort="Lu, Shan" uniqKey="Lu S" first="Shan" last="Lu">Shan Lu</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Saif, Linda J" sort="Saif, Linda J" uniqKey="Saif L" first="Linda J." last="Saif">Linda J. Saif</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><idno type="eISSN">1098-5514</idno><imprint><date when="2007">2007</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>In 2002, severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged in humans, causing a global epidemic. By phylogenetic analysis, SARS-CoV is distinct from known CoVs and most closely related to group 2 CoVs. However, no antigenic cross-reactivity between SARS-CoV and known CoVs was conclusively and consistently demonstrated except for group 1 animal CoVs. We analyzed this cross-reactivity by an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using specific antisera to animal CoVs and SARS-CoV and SARS patient convalescent-phase or negative sera. Moderate two-way cross-reactivity between SARS-CoV and porcine CoVs (transmissible gastroenteritis CoV [TGEV] and porcine respiratory CoV [PRCV]) was mediated through the N but not the spike protein, whereas weaker cross-reactivity occurred with feline (feline infectious peritonitis virus) and canine CoVs. Using <italic>Escherichia coli</italic>-expressed recombinant SARS-CoV N protein and fragments, the cross-reactive region was localized between amino acids (aa) 120 to 208. The N-protein fragments comprising aa 360 to 412 and aa 1 to 213 reacted specifically with SARS convalescent-phase sera but not with negative human sera in ELISA; the fragment comprising aa 1 to 213 cross-reacted with antisera to animal CoVs, whereas the fragment comprising aa 360 to 412 did not cross-react and could be a potential candidate for SARS diagnosis. Particularly noteworthy, a single substitution at aa 120 of PRCV N protein diminished the cross-reactivity. We also demonstrated that the cross-reactivity is not universal for all group 1 CoVs, because HCoV-NL63 did not cross-react with SARS-CoV. One-way cross-reactivity of HCoV-NL63 with group 1 CoVs was localized to aa 1 to 39 and at least one other antigenic site in the N-protein C terminus, differing from the cross-reactive region identified in SARS-CoV N protein. The observed cross-reactivity is not a consequence of a higher level of amino acid identity between SARS-CoV and porcine CoV nucleoproteins, because sequence comparisons indicated that SARS-CoV N protein has amino acid identity similar to that of infectious bronchitis virus N protein and shares a higher level of identity with bovine CoV N protein within the cross-reactive region. The TGEV and SARS-CoV N proteins are RNA chaperons with long disordered regions. We speculate that during natural infection, antibodies target similar short antigenic sites within the N proteins of SARS-CoV and porcine group 1 CoVs that are exposed to an immune response. Identification of the cross-reactive and non-cross-reactive N-protein regions allows development of SARS-CoV-specific antibody assays for screening animal and human sera.</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">J Virol</journal-id><journal-title>Journal of Virology</journal-title><issn pub-type="ppub">0022-538X</issn><issn pub-type="epub">1098-5514</issn><publisher><publisher-name>American Society for Microbiology (ASM)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">17913799</article-id><article-id pub-id-type="pmc">2168854</article-id><article-id pub-id-type="publisher-id">1169-07</article-id><article-id pub-id-type="doi">10.1128/JVI.01169-07</article-id><article-categories><subj-group subj-group-type="heading"><subject>Genetic Diversity and Evolution</subject></subj-group></article-categories><title-group><article-title>Two-Way Antigenic Cross-Reactivity between Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Group 1 Animal CoVs Is Mediated through an Antigenic Site in the N-Terminal Region of the SARS-CoV Nucleoprotein<xref ref-type="fn" rid="fn1">▿</xref> </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Vlasova</surname><given-names>Anastasia N.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Xinsheng</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Hasoksuz</surname><given-names>Mustafa</given-names></name><xref ref-type="aff" rid="aff1">2</xref></contrib><contrib contrib-type="author"><name><surname>Nagesha</surname><given-names>Hadya S.</given-names></name><xref ref-type="aff" rid="aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Haynes</surname><given-names>Lia M.</given-names></name><xref ref-type="aff" rid="aff1">3</xref></contrib><contrib contrib-type="author"><name><surname>Fang</surname><given-names>Ying</given-names></name><xref ref-type="aff" rid="aff1">4</xref></contrib><contrib contrib-type="author"><name><surname>Lu</surname><given-names>Shan</given-names></name><xref ref-type="aff" rid="aff1">5</xref></contrib><contrib contrib-type="author"><name><surname>Saif</surname><given-names>Linda J.</given-names></name><xref ref-type="aff" rid="aff1">1</xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff1">Food Animal Health Research Program, Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, Ohio 44691-4096,<label>1</label> Istanbul University, Faculty of Veterinary Medicine, Department of Virology, Avcilar 34320, Istanbul, Turkey,<label>2</label> Centers for Disease Control and Prevention, National Center for Immunization and Respiratory Diseases, Division of Viral Diseases, 1600 Clifton Road, Atlanta, Georgia 30333,<label>3</label> Center for Infectious Disease Research and Vaccinology, Veterinary Science Department, South Dakota State University, Brookings, South Dakota 57007,<label>4</label> Department of Medicine, Medical School, University of Massachusetts, 364 Plantation Street, Worcester, Massachusetts 01605<label>5</label></aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: Food Animal Health Research Program, Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development Center, The Ohio State University, 1680 Madison Avenue, Wooster, OH 44691. Phone: (330) 263-3744. Fax: (330) 263-3677. E-mail: <email>saif.2@osu.edu</email></p></fn></author-notes><pub-date pub-type="ppub"><month>12</month><year>2007</year></pub-date><pub-date pub-type="epub"><day>3</day><month>10</month><year>2007</year></pub-date><volume>81</volume><issue>24</issue><fpage>13365</fpage><lpage>13377</lpage><history><date date-type="received"><day>29</day><month>5</month><year>2007</year></date><date date-type="accepted"><day>26</day><month>9</month><year>2007</year></date></history><permissions><copyright-statement>Copyright © 2007, American Society for Microbiology</copyright-statement></permissions><self-uri xlink:title="pdf" xlink:href="zjv02407013365.pdf"></self-uri><abstract><p>In 2002, severe acute respiratory syndrome-associated coronavirus (SARS-CoV) emerged in humans, causing a global epidemic. By phylogenetic analysis, SARS-CoV is distinct from known CoVs and most closely related to group 2 CoVs. However, no antigenic cross-reactivity between SARS-CoV and known CoVs was conclusively and consistently demonstrated except for group 1 animal CoVs. We analyzed this cross-reactivity by an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis using specific antisera to animal CoVs and SARS-CoV and SARS patient convalescent-phase or negative sera. Moderate two-way cross-reactivity between SARS-CoV and porcine CoVs (transmissible gastroenteritis CoV [TGEV] and porcine respiratory CoV [PRCV]) was mediated through the N but not the spike protein, whereas weaker cross-reactivity occurred with feline (feline infectious peritonitis virus) and canine CoVs. Using <italic>Escherichia coli</italic>-expressed recombinant SARS-CoV N protein and fragments, the cross-reactive region was localized between amino acids (aa) 120 to 208. The N-protein fragments comprising aa 360 to 412 and aa 1 to 213 reacted specifically with SARS convalescent-phase sera but not with negative human sera in ELISA; the fragment comprising aa 1 to 213 cross-reacted with antisera to animal CoVs, whereas the fragment comprising aa 360 to 412 did not cross-react and could be a potential candidate for SARS diagnosis. Particularly noteworthy, a single substitution at aa 120 of PRCV N protein diminished the cross-reactivity. We also demonstrated that the cross-reactivity is not universal for all group 1 CoVs, because HCoV-NL63 did not cross-react with SARS-CoV. One-way cross-reactivity of HCoV-NL63 with group 1 CoVs was localized to aa 1 to 39 and at least one other antigenic site in the N-protein C terminus, differing from the cross-reactive region identified in SARS-CoV N protein. The observed cross-reactivity is not a consequence of a higher level of amino acid identity between SARS-CoV and porcine CoV nucleoproteins, because sequence comparisons indicated that SARS-CoV N protein has amino acid identity similar to that of infectious bronchitis virus N protein and shares a higher level of identity with bovine CoV N protein within the cross-reactive region. The TGEV and SARS-CoV N proteins are RNA chaperons with long disordered regions. We speculate that during natural infection, antibodies target similar short antigenic sites within the N proteins of SARS-CoV and porcine group 1 CoVs that are exposed to an immune response. Identification of the cross-reactive and non-cross-reactive N-protein regions allows development of SARS-CoV-specific antibody assays for screening animal and human sera.</p></abstract></article-meta></front><floats-wrap><fig position="float" id="f1"><label>FIG. 1.</label><caption><p>Schematic diagram of the eight SARS-CoV N-protein fragments (covering the whole N-protein sequence) and the full-length N protein.</p></caption><graphic xlink:href="zjv0240700460001"></graphic></fig><fig position="float" id="f2"><label>FIG. 2.</label><caption><p>Cross-reactivities of the <italic>E. coli</italic>-derived unpurified SARS-CoV N-protein fragments as assessed by Western blot analysis with SARS convalescent-phase serum (a), SARS-CoV N MAbs (b, c), or gnotobiotic pig anti-TGEV serum (d).</p></caption><graphic xlink:href="zjv0240700460002"></graphic></fig><fig position="float" id="f3"><label>FIG. 3.</label><caption><p>Cross-reactivities of the <italic>E. coli</italic>-derived, purified HCoV-NL63 N-protein fragments as assessed by Western blot analysis with guinea pig HCoV-NL63 hyperimmune antiserum (a), gnotobiotic pig PRCV-ISU1 antiserum (b), or gnotobiotic pig TGEV-P115 antiserum (c).</p></caption><graphic xlink:href="zjv0240700460003"></graphic></fig><table-wrap position="float" id="t1"><label>TABLE 1.</label><caption><p>Oligonucleotides used for recombinant protein expression, lengths of coding sequences, and predicted sizes of recombinant proteins</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Coronavirus</th><th colspan="1" rowspan="1" align="center" valign="bottom">Fragment (nt)</th><th colspan="1" rowspan="1" align="center" valign="bottom">Oligonucleotide sequences<xref ref-type="table-fn" rid="t1fn1"><italic>a</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Fragment length (nt)</th><th colspan="1" rowspan="1" align="center" valign="bottom">Predicted recombinant protein mass (kDa)</th></tr></thead><tbody><tr><td colspan="1" rowspan="2" align="left" valign="middle">TGEV (M6 and P115) and PRCV-ISU1</td><td colspan="1" rowspan="2" align="left" valign="middle">1-383</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAATGGCCAACCAGGGACAAC</td><td colspan="1" rowspan="2" align="left" valign="middle">1,149</td><td colspan="1" rowspan="2" align="left" valign="middle">43</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTCAGCGGCCGCGTTCGTTACCTCRTCAAT</td></tr><tr><td colspan="1" rowspan="8" align="left" valign="middle">HCoV-NL63</td><td colspan="1" rowspan="2" align="left" valign="middle">1-378</td><td colspan="1" rowspan="1" align="left" valign="top">F, ATGCGAATTCGATGGCTAGTGTAAATTGG</td><td colspan="1" rowspan="2" align="left" valign="middle">1,134</td><td colspan="1" rowspan="2" align="left" valign="middle">44</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GCATGCGGCCGCATGCAAAACCTCGTTG</td></tr><tr><td colspan="1" rowspan="2" align="left" valign="middle">1-183</td><td colspan="1" rowspan="1" align="left" valign="top">F, ATGCGAATTCGATGGCTAGTGTAAATTGG</td><td colspan="1" rowspan="2" align="left" valign="middle">549</td><td colspan="1" rowspan="2" align="left" valign="middle">21</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GCATGCGGCCGCGTTAGAATCAGAACGA</td></tr><tr><td colspan="1" rowspan="2" align="left" valign="middle">182-378</td><td colspan="1" rowspan="1" align="left" valign="top">F, ATGCGAATTCGTCTAACCAGTCTTCTTC</td><td colspan="1" rowspan="2" align="left" valign="middle">588</td><td colspan="1" rowspan="2" align="left" valign="middle">23</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GCATGCGGCCGCATGCAAAACCTCGTTG</td></tr><tr><td colspan="1" rowspan="2" align="left" valign="middle">39-183</td><td colspan="1" rowspan="1" align="left" valign="middle">F, ATGCGAATTCGAAGGACCTTAAATTCAG</td><td colspan="1" rowspan="2" align="left" valign="middle">432</td><td colspan="1" rowspan="2" align="left" valign="middle">16.8</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GCATGCGGCCGCGTTAGAATCAGAACGA</td></tr><tr><td colspan="1" rowspan="18" align="left" valign="middle">SARS-CoV</td><td colspan="1" rowspan="2" align="left" valign="middle">1-72</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAATGTCTGATAATGGACC</td><td colspan="1" rowspan="2" align="left" valign="middle">216</td><td colspan="1" rowspan="2" align="left" valign="middle">7.8</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACGCCCTGGCCTCGAGGG</td></tr><tr><td colspan="1" rowspan="2" align="center" valign="middle">120-208</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAGCTTCACTTCCCTACGG</td><td colspan="1" rowspan="2" align="center" valign="middle">264</td><td colspan="1" rowspan="2" align="center" valign="middle">9.7</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACAGGAGAATTTCCCCTAC</td></tr><tr><td colspan="1" rowspan="2" align="left" valign="middle">212-349</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAGCTAGCGGAGGTGGTGA</td><td colspan="1" rowspan="2" align="left" valign="middle">411</td><td colspan="1" rowspan="2" align="left" valign="middle">15</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACGTCTTTGAATTGTGGATC</td></tr><tr><td colspan="1" rowspan="2" align="left" valign="middle">360-412</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAGCATACAAAACATTCCC</td><td colspan="1" rowspan="2" align="left" valign="middle">156</td><td colspan="1" rowspan="2" align="left" valign="middle">5.7</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="middle">R, GTGAGTCGACCATGGAATTTTGAAGTTG</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"></td><td colspan="1" rowspan="2" align="left" valign="middle">70-213</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAGGCCAGGGCGTTCCAATC</td><td colspan="1" rowspan="2" align="left" valign="middle">429</td><td colspan="1" rowspan="2" align="left" valign="middle">15.7</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACGCTAGCCATTCGAGCAGG</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"></td><td colspan="1" rowspan="2" align="left" valign="middle">337-422</td><td colspan="1" rowspan="1" align="left" valign="middle">F, TGACGAATTCAGCCATTAAATTGGATGA</td><td colspan="1" rowspan="2" align="left" valign="middle">255</td><td colspan="1" rowspan="2" align="left" valign="middle">9.4</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACTGCCTGAGTTGAATCAG</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"></td><td colspan="1" rowspan="2" align="left" valign="middle">1-213</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAATGTCTGATAATGGACC</td><td colspan="1" rowspan="2" align="left" valign="middle">639</td><td colspan="1" rowspan="2" align="left" valign="middle">23</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACGCTAGCCATTCGAGCAGG</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"></td><td colspan="1" rowspan="2" align="left" valign="middle">212-422</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAGCTAGCGGAGGTGGTGA</td><td colspan="1" rowspan="2" align="left" valign="middle">630</td><td colspan="1" rowspan="2" align="left" valign="middle">23</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACTGCCTGAGTTGAATCAG</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"></td><td colspan="1" rowspan="2" align="left" valign="middle">1-422</td><td colspan="1" rowspan="1" align="left" valign="top">F, TGACGAATTCAATGTCTGATAATGGACC</td><td colspan="1" rowspan="2" align="left" valign="middle">1,266</td><td colspan="1" rowspan="2" align="left" valign="middle">49</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">R, GTGAGTCGACTGCCTGAGTTGAATCAG</td></tr></tbody></table><table-wrap-foot><fn id="t1fn1"><label>a</label><p>F, forward; R, reverse.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t2"><label>TABLE 2.</label><caption><p>ELISA results for testing intergroup cross-reactivities of CoVs by using unpurified group 1 to 3 CoVs (as CoV-infected crude cell lysates) and specific hyperimmune antisera</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="3" align="center" valign="middle">Antiserum (host)<xref ref-type="table-fn" rid="t2fn2"><italic>b</italic></xref></th><th colspan="9" rowspan="1" align="center" valign="bottom">ELISA titer (<italic>R</italic>%)<xref ref-type="table-fn" rid="t2fn1"><italic>a</italic></xref> for indicated coronavirus
<hr></hr></th></tr><tr><th colspan="6" rowspan="1" align="center" valign="bottom">Group 1
<hr></hr></th><th colspan="1" rowspan="1" align="center" valign="bottom">Group 2a
<hr></hr></th><th colspan="1" rowspan="1" align="center" valign="bottom">Group 2b
<hr></hr></th><th colspan="1" rowspan="1" align="center" valign="bottom">Group 3
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">TGEV-M6<xref ref-type="table-fn" rid="t2fn3"><italic>c</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">TGEV-P115</th><th colspan="1" rowspan="1" align="center" valign="bottom">PRCV-ISU1</th><th colspan="1" rowspan="1" align="center" valign="bottom">CCoV-UCD1</th><th colspan="1" rowspan="1" align="center" valign="bottom">FIPV 79-1146</th><th colspan="1" rowspan="1" align="center" valign="bottom">HCoV-NL63</th><th colspan="1" rowspan="1" align="center" valign="bottom">BCoV-Mebus<xref ref-type="table-fn" rid="t2fn4"><italic>d</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">SARS-CoV Urbani</th><th colspan="1" rowspan="1" align="center" valign="bottom">IBV-Massachusetts<xref ref-type="table-fn" rid="t2fn4"><italic>d</italic></xref></th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 1</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-M6 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>6,400 (100)</bold><xref ref-type="table-fn" rid="t2fn7"><italic>g</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400 (100)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600 (50)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="center" valign="top">Negative<xref ref-type="table-fn" rid="t2fn5"><italic>e</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">100 (1.1)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-P115 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200 (100)</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>51,200 (100)</bold></td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (50)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800 (1.1)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> PRCV-ISU1 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">102,400 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">102,400 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>102,400 (100)</bold></td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800 (0.55)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CCoV-UCD1 (GP)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (50)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (50)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (70)</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>12,800 (100)</bold></td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800 (100)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> FIPV 79-1146 (GP)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200 (35)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400 (100)</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>6,400 (100)</bold></td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> HCoV-NL63 (GP)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>12,800 (100)</bold></td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2a</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> BCoV-Mebus (GP/Gn calf)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>25,600 (100)</bold></td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2b</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> SARS-CoV-Urbani (mouse)<xref ref-type="table-fn" rid="t2fn6"><italic>f</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">200 (1.1)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200 (1.1)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100 (0.55)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>25,600 (100)</bold></td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 3</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> IBV-Massachusetts (GP/chicken)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top"><bold>51,200,102,400 (100)</bold></td></tr></tbody></table><table-wrap-foot><fn id="t2fn1"><label>a</label><p><italic>R</italic>% values are indicated in parentheses in cases where they were possible to calculate, as follows: <inline-formula><tex-math id="M1">\documentclass[10pt]{article}
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\begin{equation*}R\%=100(\sqrt{r_{1}{\times}r_{2}}\end{equation*}\end{document}</tex-math></inline-formula>), where <italic>r</italic><sub>1</sub> is the heterologous titer of strain 2 divided by the homologous titer of strain 1, and <italic>r</italic><sub>2</sub> is the heterologous titer of strain 1 divided by the homologous titer of strain 1.</p></fn><fn id="t2fn2"><label>b</label><p>Negative gnotobiotic (Gn) pig, Gn calf, guinea pig (GP), human (CoV samples 9 to 24), and mouse (Sigma) sera were used as negative controls.</p></fn><fn id="t2fn3"><label>c</label><p>Mock-infected host cell lines HRT-18, ST, Vero E6, CrFK, A72, and LLCK12 and negative chicken embryo allantoic fluids were used as negative controls.</p></fn><fn id="t2fn4"><label>d</label><p>Only one representative each from groups 2a and 3 is given because the results were the same for the other tested CoV strains from these groups.</p></fn><fn id="t2fn5"><label>e</label><p>Negative indicates an ELISA titer of <100.</p></fn><fn id="t2fn6"><label>f</label><p>Mouse hyperimmune antiserum to inactivated SARS-CoV-Urbani was provided by CDC (Atlanta, GA).</p></fn><fn id="t2fn7"><label>g</label><p>Boldface values represent homologous titers for each CoV.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t3"><label>TABLE 3.</label><caption><p>Assessment by ELISA of homologous reactivity and cross-reactivity of the purified recombinant N proteins produced in <italic>E. coli</italic> (SARS-CoV N-protein fragments, HCoV-NL63 N protein, and TGEV-M6, TGEV-P115, and PRCV N proteins) and unpurified SARS-CoV S protein produced in human 293T cells</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="4" align="center" valign="middle">Antiserum (host)<xref ref-type="table-fn" rid="t3fn3"><italic>c</italic></xref>,<xref ref-type="table-fn" rid="t3fn4"><italic>d</italic></xref></th><th colspan="14" rowspan="1" align="center" valign="bottom">ELISA titer for indicated protein fragment
<hr></hr></th></tr><tr><th colspan="9" rowspan="2" align="center" valign="bottom">SARS-CoV N protein<xref ref-type="table-fn" rid="t3fn1"><italic>a</italic></xref><hr></hr></th><th colspan="1" rowspan="2" align="center" valign="middle">SARS-CoV S protein<xref ref-type="table-fn" rid="t3fn2"><italic>b</italic></xref></th><th colspan="4" rowspan="1" align="center" valign="bottom">Recombinant N proteins for<xref ref-type="table-fn" rid="t3fn1"><italic>a</italic></xref>:
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">TGEV-M6</th><th colspan="1" rowspan="1" align="center" valign="bottom">TGEV-P115</th><th colspan="1" rowspan="1" align="center" valign="bottom">PRCV-ISU1</th><th colspan="1" rowspan="1" align="center" valign="bottom">HCoV-NL63</th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-72</th><th colspan="1" rowspan="1" align="center" valign="bottom"><bold>aa 70-213</bold></th><th colspan="1" rowspan="1" align="center" valign="bottom"><bold>aa 120-208</bold></th><th colspan="1" rowspan="1" align="center" valign="bottom"><bold>aa 1-213<xref ref-type="table-fn" rid="t3fn5"><italic>e</italic></xref></bold></th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 212-349</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 337-422</th><th colspan="1" rowspan="1" align="center" valign="bottom"><bold>aa 360-412<xref ref-type="table-fn" rid="t3fn5"><italic>e</italic></xref></bold></th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 212-422</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-422</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-1190</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-383</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-383</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-383</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-378</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 1</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-M6 (Gn pig)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative<xref ref-type="table-fn" rid="t3fn6"><italic>f</italic></xref></td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-P115 (Gn pig)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> PRCV-ISU1 (Gn pig)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CCoV-UCD1 (GP)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> FIPV 79-1146 (GP)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> HCoV-NL63 (GP)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2a</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> BCoV-Mebus (GP/Gn calf)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2b</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> SARS-CoV-Urbani (mouse)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 3</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> IBV-Massachusetts (GP/chicken)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Human SARS convalescent-phase sera</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 3</td><td colspan="1" rowspan="1" align="char" char="." valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 4</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 5</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 6</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 7</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 8</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Human negative sera</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV samples 9-19</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">400-6,400</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 20</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV sample 21</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CoV samples 22-24</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">800-6,400</td></tr></tbody></table><table-wrap-foot><fn id="t3fn1"><label>a</label><p>HIS-Select nickel affinity gel-purified, <italic>E. coli</italic>-derived proteins from mock-transformed <italic>E. coli</italic> were used as a negative control for the ELISAs with SARS-CoV, TGEV, and PRCV N proteins.</p></fn><fn id="t3fn2"><label>b</label><p>Mock-transfected 293T cell lysate was used as a negative control for the ELISAs with the SARS-CoV S protein produced in 293T cells.</p></fn><fn id="t3fn3"><label>c</label><p>Negative gnotobiotic (Gn) pig, Gn calf, guinea pig (GP), and mouse sera were used in all ELISAs as negative controls.</p></fn><fn id="t3fn4"><label>d</label><p>SARS-CoV and TGEV N MAbs were also tested, but no cross-reactivity was shown (data not shown).</p></fn><fn id="t3fn5"><label>e</label><p>Fragments 1 to 213 and 360 to 412 contained the most antigenic sites, but only fragments 70 to 213, 120 to 208, and 1 to 213 were cross-reactive with Abs to group 1 CoVs (except NL63) and SARS-CoV.</p></fn><fn id="t3fn6"><label>f</label><p>Negative is defined as an ELISA titer of <100.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t4"><label>TABLE 4.</label><caption><p>Assessment by ELISA of homologous reactivity and cross-reactivity of the HCoV-NL63 recombinant N-protein fragments</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Antiserum or TGEV anti-N MAb (host)<xref ref-type="table-fn" rid="t4fn2"><italic>b</italic></xref></th><th colspan="4" rowspan="1" align="center" valign="bottom">ELISA titer for indicated HCoV-NL63 N-protein fragment<xref ref-type="table-fn" rid="t4fn1"><italic>a</italic></xref><hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-183</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 182-378</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 39-183</th><th colspan="1" rowspan="1" align="center" valign="bottom">aa 1-378</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 1</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-M6 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="center" valign="top">Negative<xref ref-type="table-fn" rid="t4fn3"><italic>c</italic></xref></td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> TGEV-P115 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> PRCV-ISU1 (Gn pig)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">12,800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> CCoV-UCD1 (GP)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> FIPV 79-1146 (GP)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">100</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> HCoV-NL63 (GP)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="char" char="." valign="top">51,200</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> 25H7</td><td colspan="1" rowspan="1" align="char" char="." valign="top">800</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> 14E3</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6,400</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> 14G9</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1,600</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3,200</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2a</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> BCoV-Mebus (GP/Gn calf)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 2b</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> SARS-CoV-Urbani (mouse)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">Group 3</td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="center" valign="top"></td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top"> IBV-Massachusetts (GP/chicken)</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td><td colspan="1" rowspan="1" align="center" valign="top">Negative</td></tr></tbody></table><table-wrap-foot><fn id="t4fn1"><label>a</label><p>HIS-Select nickel affinity gel-purified, <italic>E. coli</italic>-derived proteins from mock-transformed <italic>E. coli</italic> were used as a negative control for the ELISAs with HCoV-NL63 N-protein fragments.</p></fn><fn id="t4fn2"><label>b</label><p>Negative gnotobiotic (Gn), guinea pig (GP), and mouse sera and negative mouse ascites (SP2/0) were used in all ELISAs as negative control sera. 25H7, 14E3, and 14G9 are TGEV-M6 anti-N MAbs.</p></fn><fn id="t4fn3"><label>c</label><p>Negative is defined as an ELISA titer of <100.</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t5"><label>TABLE 5.</label><caption><p>Amino acid identities between SARS-CoV-Urbani N protein and N proteins of the other CoVs</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Sequence (GenBank accession no.)</th><th colspan="2" rowspan="1" align="center" valign="bottom">% SARS-CoV N-protein aa identity
<hr></hr></th><th colspan="1" rowspan="2" align="center" valign="middle">Cross-reactivity</th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Total</th><th colspan="1" rowspan="1" align="center" valign="bottom">Within cross-reactive region (aa 70-213)</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="left" valign="top">TGEV-M6 (DQ811785)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25.8</td><td colspan="1" rowspan="1" align="char" char="." valign="top">36</td><td colspan="1" rowspan="1" align="center" valign="top">Yes</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">TGEV-P115 (DQ811788)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25.8</td><td colspan="1" rowspan="1" align="char" char="." valign="top">36</td><td colspan="1" rowspan="1" align="center" valign="top">Yes</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">PRCV-ISU1 (DQ811787)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">25.1</td><td colspan="1" rowspan="1" align="char" char="." valign="top">35.6</td><td colspan="1" rowspan="1" align="center" valign="top">Yes</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">CCoV-BGF10 (AY342160)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">27</td><td colspan="1" rowspan="1" align="char" char="." valign="top">40.5</td><td colspan="1" rowspan="1" align="center" valign="top">Yes</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">FIPV-WSU 79/1146 (NC_007025)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">26</td><td colspan="1" rowspan="1" align="char" char="." valign="top">35.6</td><td colspan="1" rowspan="1" align="center" valign="top">Yes</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">HCoV-NL63 (DQ846901)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">27</td><td colspan="1" rowspan="1" align="char" char="." valign="top">35.6</td><td colspan="1" rowspan="1" align="center" valign="top">No</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">BCoV-Mebus (U00735)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">34.6</td><td colspan="1" rowspan="1" align="char" char="." valign="top">46.8</td><td colspan="1" rowspan="1" align="center" valign="top">No</td></tr><tr><td colspan="1" rowspan="1" align="left" valign="top">IBV-Massachusetts 41 (AY851295)</td><td colspan="1" rowspan="1" align="char" char="." valign="top">26</td><td colspan="1" rowspan="1" align="char" char="." valign="top">35.6</td><td colspan="1" rowspan="1" align="center" valign="top">No</td></tr></tbody></table></table-wrap></floats-wrap></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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