A rapid point of care immunoswab assay for SARS-CoV detection
Identifieur interne : 000E56 ( Pmc/Checkpoint ); précédent : 000E55; suivant : 000E57A rapid point of care immunoswab assay for SARS-CoV detection
Auteurs : Sriram Kammila [Canada] ; Dipankar Das [Canada] ; Pravin K. Bhatnagar [Canada] ; Hoon H. Sunwoo [Canada] ; Gustavo Zayas-Zamora [Canada] ; Malcolm King [Canada] ; Mavanur R. Suresh [Canada]Source :
- Journal of Virological Methods [ 0166-0934 ] ; 2008.
Abstract
The emergence of severe acute respiratory syndrome (SARS) resulted in several outbreaks worldwide. Early tests for diagnosis were not always conclusive in identifying a SARS suspected patient. Nucleocapsid protein (NP) is the most predominant virus derived structural protein which is shed in high amounts in serum and nasopharyngeal aspirate during the first week of infection. As part of such efforts, a simple, easy to use immunoswab method was developed by generating a panel of monoclonal antibodies (MAbs), Bispecific MAbs and chicken polyclonal IgY antibody against the SARS-CoV nucleocapsid protein (NP). Employing the MAb-based immunoswab, an NP concentration of 200 pg/mL in saline and pig nasopharyngeal aspirate, and 500 pg/mL in rabbit serum were detected. BsMAb-based immunoswabs detected an NP concentration of 20 pg/mL in saline, 500 pg/mL in rabbit serum and 20–200 pg/mL in pig nasopharyngeal aspirate. Polyclonal IgY-based immunoswabs detected an NP concentration of 10 pg/mL in pig nasopharyngeal aspirate providing the most sensitive SARS point of care assay. Results show that the robust immunoswab method of detecting SARS-CoV NP antigen can be developed into an easy and effective way of identifying SARS suspected individuals during a future SARS epidemic, thereby reducing and containing the transmission. The key feature of this simple immunoswab diagnostic assay is its ability to detect the presence of the SARS-CoV antigen within 45–60 min with the availability of the body fluid samples.
Url:
DOI: 10.1016/j.jviromet.2008.05.023
PubMed: 18620761
PubMed Central: 2678951
Affiliations:
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<front><div type="abstract" xml:lang="en"><p>The emergence of severe acute respiratory syndrome (SARS) resulted in several outbreaks worldwide. Early tests for diagnosis were not always conclusive in identifying a SARS suspected patient. Nucleocapsid protein (NP) is the most predominant virus derived structural protein which is shed in high amounts in serum and nasopharyngeal aspirate during the first week of infection. As part of such efforts, a simple, easy to use immunoswab method was developed by generating a panel of monoclonal antibodies (MAbs), Bispecific MAbs and chicken polyclonal IgY antibody against the SARS-CoV nucleocapsid protein (NP). Employing the MAb-based immunoswab, an NP concentration of 200 pg/mL in saline and pig nasopharyngeal aspirate, and 500 pg/mL in rabbit serum were detected. BsMAb-based immunoswabs detected an NP concentration of 20 pg/mL in saline, 500 pg/mL in rabbit serum and 20–200 pg/mL in pig nasopharyngeal aspirate. Polyclonal IgY-based immunoswabs detected an NP concentration of 10 pg/mL in pig nasopharyngeal aspirate providing the most sensitive SARS point of care assay. Results show that the robust immunoswab method of detecting SARS-CoV NP antigen can be developed into an easy and effective way of identifying SARS suspected individuals during a future SARS epidemic, thereby reducing and containing the transmission. The key feature of this simple immunoswab diagnostic assay is its ability to detect the presence of the SARS-CoV antigen within 45–60 min with the availability of the body fluid samples.</p>
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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol Methods</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Virol. Methods</journal-id>
<journal-title-group><journal-title>Journal of Virological Methods</journal-title>
</journal-title-group>
<issn pub-type="ppub">0166-0934</issn>
<issn pub-type="epub">1879-0984</issn>
<publisher><publisher-name>Elsevier B.V.</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">18620761</article-id>
<article-id pub-id-type="pmc">2678951</article-id>
<article-id pub-id-type="publisher-id">S0166-0934(08)00167-5</article-id>
<article-id pub-id-type="doi">10.1016/j.jviromet.2008.05.023</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>A rapid point of care immunoswab assay for SARS-CoV detection</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Kammila</surname>
<given-names>Sriram</given-names>
</name>
<xref rid="aff1" ref-type="aff">a</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Das</surname>
<given-names>Dipankar</given-names>
</name>
<xref rid="aff1" ref-type="aff">a</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Bhatnagar</surname>
<given-names>Pravin K.</given-names>
</name>
<xref rid="aff1" ref-type="aff">a</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Sunwoo</surname>
<given-names>Hoon H.</given-names>
</name>
<xref rid="aff1" ref-type="aff">a</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Zayas-Zamora</surname>
<given-names>Gustavo</given-names>
</name>
<xref rid="aff2" ref-type="aff">b</xref>
</contrib>
<contrib contrib-type="author"><name><surname>King</surname>
<given-names>Malcolm</given-names>
</name>
<xref rid="aff2" ref-type="aff">b</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Suresh</surname>
<given-names>Mavanur R.</given-names>
</name>
<email>msuresh@pharmacy.ualberta.ca</email>
<xref rid="aff1" ref-type="aff">a</xref>
<xref rid="cor1" ref-type="corresp">⁎</xref>
</contrib>
</contrib-group>
<aff id="aff1"><label>a</label>
Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, 11304-89 Avenue, Edmonton, Alberta, Canada T6G 2N8</aff>
<aff id="aff2"><label>b</label>
Mucophysiology Laboratory, 173 Heritage Medical Research Centre, Department of Medicine, Faculty of Medicine & Dentistry, University of Alberta, Alberta, Canada T6G 2S2</aff>
<author-notes><corresp id="cor1"><label>⁎</label>
Corresponding author. Tel.: +1 780 492 9233; fax: +1 780 492 1217. <email>msuresh@pharmacy.ualberta.ca</email>
</corresp>
</author-notes>
<pub-date pub-type="pmc-release"><day>11</day>
<month>7</month>
<year>2008</year>
</pub-date>
<pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>9</month>
<year>2008</year>
</pub-date>
<pub-date pub-type="epub"><day>11</day>
<month>7</month>
<year>2008</year>
</pub-date>
<volume>152</volume>
<issue>1</issue>
<fpage>77</fpage>
<lpage>84</lpage>
<history><date date-type="received"><day>17</day>
<month>1</month>
<year>2008</year>
</date>
<date date-type="rev-recd"><day>18</day>
<month>4</month>
<year>2008</year>
</date>
<date date-type="accepted"><day>8</day>
<month>5</month>
<year>2008</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2008 Elsevier B.V. All rights reserved.</copyright-statement>
<copyright-year>2008</copyright-year>
<copyright-holder>Elsevier B.V.</copyright-holder>
<license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p>
</license>
</permissions>
<abstract><p>The emergence of severe acute respiratory syndrome (SARS) resulted in several outbreaks worldwide. Early tests for diagnosis were not always conclusive in identifying a SARS suspected patient. Nucleocapsid protein (NP) is the most predominant virus derived structural protein which is shed in high amounts in serum and nasopharyngeal aspirate during the first week of infection. As part of such efforts, a simple, easy to use immunoswab method was developed by generating a panel of monoclonal antibodies (MAbs), Bispecific MAbs and chicken polyclonal IgY antibody against the SARS-CoV nucleocapsid protein (NP). Employing the MAb-based immunoswab, an NP concentration of 200 pg/mL in saline and pig nasopharyngeal aspirate, and 500 pg/mL in rabbit serum were detected. BsMAb-based immunoswabs detected an NP concentration of 20 pg/mL in saline, 500 pg/mL in rabbit serum and 20–200 pg/mL in pig nasopharyngeal aspirate. Polyclonal IgY-based immunoswabs detected an NP concentration of 10 pg/mL in pig nasopharyngeal aspirate providing the most sensitive SARS point of care assay. Results show that the robust immunoswab method of detecting SARS-CoV NP antigen can be developed into an easy and effective way of identifying SARS suspected individuals during a future SARS epidemic, thereby reducing and containing the transmission. The key feature of this simple immunoswab diagnostic assay is its ability to detect the presence of the SARS-CoV antigen within 45–60 min with the availability of the body fluid samples.</p>
</abstract>
<kwd-group><title>Keywords</title>
<kwd>Bispecific monoclonal antibody</kwd>
<kwd>SARS-CoV</kwd>
<kwd>Nucleocapsid protein</kwd>
<kwd>Immunoswab assay</kwd>
</kwd-group>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>Canada</li>
</country>
</list>
<tree><country name="Canada"><noRegion><name sortKey="Kammila, Sriram" sort="Kammila, Sriram" uniqKey="Kammila S" first="Sriram" last="Kammila">Sriram Kammila</name>
</noRegion>
<name sortKey="Bhatnagar, Pravin K" sort="Bhatnagar, Pravin K" uniqKey="Bhatnagar P" first="Pravin K." last="Bhatnagar">Pravin K. Bhatnagar</name>
<name sortKey="Das, Dipankar" sort="Das, Dipankar" uniqKey="Das D" first="Dipankar" last="Das">Dipankar Das</name>
<name sortKey="King, Malcolm" sort="King, Malcolm" uniqKey="King M" first="Malcolm" last="King">Malcolm King</name>
<name sortKey="Sunwoo, Hoon H" sort="Sunwoo, Hoon H" uniqKey="Sunwoo H" first="Hoon H." last="Sunwoo">Hoon H. Sunwoo</name>
<name sortKey="Suresh, Mavanur R" sort="Suresh, Mavanur R" uniqKey="Suresh M" first="Mavanur R." last="Suresh">Mavanur R. Suresh</name>
<name sortKey="Zayas Zamora, Gustavo" sort="Zayas Zamora, Gustavo" uniqKey="Zayas Zamora G" first="Gustavo" last="Zayas-Zamora">Gustavo Zayas-Zamora</name>
</country>
</tree>
</affiliations>
</record>
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