Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)
Identifieur interne : 005221 ( Main/Merge ); précédent : 005220; suivant : 005222Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)
Auteurs : SHIXIA WANG [États-Unis] ; Pavlo Sakhatskyy [États-Unis] ; Te-Hui W. Chou [États-Unis] ; SHAN LU [États-Unis]Source :
- Journal of immunological methods [ 0022-1759 ] ; 2005.
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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.
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Chou</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Shan Lu" sort="Shan Lu" uniqKey="Shan Lu" last="Shan Lu">SHAN LU</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">05-0333869</idno><date when="2005">2005</date><idno type="stanalyst">PASCAL 05-0333869 INIST</idno><idno type="RBID">Pascal:05-0333869</idno><idno type="wicri:Area/PascalFrancis/Corpus">000637</idno><idno type="wicri:Area/PascalFrancis/Curation">000353</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000701</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000701</idno><idno type="wicri:doubleKey">0022-1759:2005:Shixia Wang:assays:for:the</idno><idno type="wicri:Area/Main/Merge">005221</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)</title><author><name sortKey="Shixia Wang" sort="Shixia Wang" uniqKey="Shixia Wang" last="Shixia Wang">SHIXIA WANG</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Sakhatskyy, Pavlo" sort="Sakhatskyy, Pavlo" uniqKey="Sakhatskyy P" first="Pavlo" last="Sakhatskyy">Pavlo Sakhatskyy</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></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><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Shan Lu" sort="Shan Lu" uniqKey="Shan Lu" last="Shan Lu">SHAN LU</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Laboratory of Nucleic Acid Vaccines, Department of Medicine, University of Massachusetts Medical School, 364 Plantation Street, Lazare Research Building</s1><s2>Worcester, MA 01605-2397</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Massachusetts</region></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of immunological methods</title><title level="j" type="abbreviated">J. immunol. methods</title><idno type="ISSN">0022-1759</idno><imprint><date when="2005">2005</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of immunological methods</title><title level="j" type="abbreviated">J. immunol. methods</title><idno type="ISSN">0022-1759</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Immunological method</term><term>Neutralization</term><term>Neutralizing antibody</term><term>Severe acute respiratory syndrome</term><term>Severe acute respiratory syndrome virus</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Virus syndrome respiratoire aigu sévère</term><term>Anticorps neutralisant</term><term>Neutralisation</term><term>Méthode immunologique</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><country name="États-Unis"><region name="Massachusetts"><name sortKey="Shixia Wang" sort="Shixia Wang" uniqKey="Shixia Wang" last="Shixia Wang">SHIXIA WANG</name></region><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="Sakhatskyy, Pavlo" sort="Sakhatskyy, Pavlo" uniqKey="Sakhatskyy P" first="Pavlo" last="Sakhatskyy">Pavlo Sakhatskyy</name><name sortKey="Shan Lu" sort="Shan Lu" uniqKey="Shan Lu" last="Shan Lu">SHAN LU</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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