Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)
Identifieur interne : 001388 ( Pmc/Checkpoint ); précédent : 001387; suivant : 001389Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)
Auteurs : Shixia Wang ; Pavlo Sakhatskyy ; Te-Hui W. Chou ; Shan LuSource :
- Journal of Immunological Methods [ 0022-1759 ] ; 2005.
Abstract
Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A540. The neutralizing antibody titers can be easily determined with either of the two new assays. In this report, we described the utility of these two new neutralization assays in measuring the neutralizing activities against SCV infection from rabbit sera immunized with various forms of spike protein of SCV.
Url:
DOI: 10.1016/j.jim.2005.03.008
PubMed: 15894326
PubMed Central: 7094753
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)</title><author><name sortKey="Wang, Shixia" sort="Wang, Shixia" uniqKey="Wang S" first="Shixia" last="Wang">Shixia Wang</name></author><author><name sortKey="Sakhatskyy, Pavlo" sort="Sakhatskyy, Pavlo" uniqKey="Sakhatskyy P" first="Pavlo" last="Sakhatskyy">Pavlo Sakhatskyy</name></author><author><name sortKey="Chou, Te Hui W" sort="Chou, Te Hui W" uniqKey="Chou T" first="Te-Hui W." last="Chou">Te-Hui W. Chou</name></author><author><name sortKey="Lu, Shan" sort="Lu, Shan" uniqKey="Lu S" first="Shan" last="Lu">Shan Lu</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">15894326</idno><idno type="pmc">7094753</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7094753</idno><idno type="RBID">PMC:7094753</idno><idno type="doi">10.1016/j.jim.2005.03.008</idno><date when="2005">2005</date><idno type="wicri:Area/Pmc/Corpus">001160</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001160</idno><idno type="wicri:Area/Pmc/Curation">001160</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001160</idno><idno type="wicri:Area/Pmc/Checkpoint">001388</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001388</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)</title><author><name sortKey="Wang, Shixia" sort="Wang, Shixia" uniqKey="Wang S" first="Shixia" last="Wang">Shixia Wang</name></author><author><name sortKey="Sakhatskyy, Pavlo" sort="Sakhatskyy, Pavlo" uniqKey="Sakhatskyy P" first="Pavlo" last="Sakhatskyy">Pavlo Sakhatskyy</name></author><author><name sortKey="Chou, Te Hui W" sort="Chou, Te Hui W" uniqKey="Chou T" first="Te-Hui W." last="Chou">Te-Hui W. Chou</name></author><author><name sortKey="Lu, Shan" sort="Lu, Shan" uniqKey="Lu S" first="Shan" last="Lu">Shan Lu</name></author></analytic><series><title level="j">Journal of Immunological Methods</title><idno type="ISSN">0022-1759</idno><idno type="eISSN">1872-7905</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A<sub>540</sub>. The neutralizing antibody titers can be easily determined with either of the two new assays. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Immunol Methods</journal-id><journal-id journal-id-type="iso-abbrev">J. Immunol. Methods</journal-id><journal-title-group><journal-title>Journal of Immunological Methods</journal-title></journal-title-group><issn pub-type="ppub">0022-1759</issn><issn pub-type="epub">1872-7905</issn><publisher><publisher-name>Elsevier B.V.</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">15894326</article-id><article-id pub-id-type="pmc">7094753</article-id><article-id pub-id-type="publisher-id">S0022-1759(05)00083-9</article-id><article-id pub-id-type="doi">10.1016/j.jim.2005.03.008</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Shixia</given-names></name></contrib><contrib contrib-type="author"><name><surname>Sakhatskyy</surname><given-names>Pavlo</given-names></name></contrib><contrib contrib-type="author"><name><surname>Chou</surname><given-names>Te-Hui W.</given-names></name></contrib><contrib contrib-type="author"><name><surname>Lu</surname><given-names>Shan</given-names></name><email>shan.lu@umassmed.edu</email><xref rid="cor1" ref-type="corresp">⁎</xref></contrib></contrib-group><aff>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building, Worcester, MA 01605-2397, United States</aff><author-notes><corresp id="cor1"><label>⁎</label>Corresponding author. Tel.: +1 508 856 6791; fax: +1 508 856 6751. <email>shan.lu@umassmed.edu</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>25</day><month>4</month><year>2005</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>6</month><year>2005</year></pub-date><pub-date pub-type="epub"><day>25</day><month>4</month><year>2005</year></pub-date><volume>301</volume><issue>1</issue><fpage>21</fpage><lpage>30</lpage><history><date date-type="received"><day>11</day><month>1</month><year>2005</year></date><date date-type="accepted"><day>6</day><month>3</month><year>2005</year></date></history><permissions><copyright-statement>Copyright © 2005 Elsevier B.V. All rights reserved.</copyright-statement><copyright-year>2005</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>Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A<sub>540</sub>. The neutralizing antibody titers can be easily determined with either of the two new assays. In this report, we described the utility of these two new neutralization assays in measuring the neutralizing activities against SCV infection from rabbit sera immunized with various forms of spike protein of SCV.</p></abstract><kwd-group><title>Abbreviations</title><kwd>SARS, Severe Acute Respiratory Syndrome</kwd><kwd>SCV, SARS associated coronavirus</kwd><kwd>CPE, cytopathic effect</kwd><kwd>PR, plaque reduction</kwd><kwd>NRS, neutral red staining</kwd><kwd>TCID<sub>50</sub>, 50% tissue culture infectious dose</kwd><kwd>HIV-1, human immunodeficiency virus type 1</kwd><kwd>MOI, multiplicity of infection</kwd><kwd>DMEM, Dulbecco modified Eagle medium</kwd></kwd-group><kwd-group><title>Keywords</title><kwd>Severe Acute Respiratory Syndrome (SARS)</kwd><kwd>Coronavirus</kwd><kwd>Neutralization assay</kwd><kwd>Anti-viral antibody</kwd></kwd-group></article-meta></front></pmc><affiliations><list></list><tree><noCountry><name sortKey="Chou, Te Hui W" sort="Chou, Te Hui W" uniqKey="Chou T" first="Te-Hui W." last="Chou">Te-Hui W. Chou</name><name sortKey="Lu, Shan" sort="Lu, Shan" uniqKey="Lu S" first="Shan" last="Lu">Shan Lu</name><name sortKey="Sakhatskyy, Pavlo" sort="Sakhatskyy, Pavlo" uniqKey="Sakhatskyy P" first="Pavlo" last="Sakhatskyy">Pavlo Sakhatskyy</name><name sortKey="Wang, Shixia" sort="Wang, Shixia" uniqKey="Wang S" first="Shixia" last="Wang">Shixia Wang</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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