Assays for the assessment of neutralizing antibody activities against Severe Acute Respiratory Syndrome (SARS) associated coronavirus (SCV)
Identifieur interne : 004920 ( Main/Exploration ); précédent : 004919; suivant : 004921Assays 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.
Descripteurs français
- KwdFr :
- MESH :
- analyse : Anticorps antiviraux.
- immunologie : Anticorps antiviraux, Syndrome respiratoire aigu sévère, Virus du SRAS.
- virologie : Syndrome respiratoire aigu sévère.
- Animaux, Cellules Vero, Reproductibilité des résultats, Tests de neutralisation.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Antibodies, Viral.
- chemical , immunology : Antibodies, Viral.
- immunology : SARS Virus, Severe Acute Respiratory Syndrome.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Chlorocebus aethiops, Neutralization Tests, Reproducibility of Results, Vero Cells.
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
Affiliations:
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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><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Antibodies, Viral (analysis)</term><term>Antibodies, Viral (immunology)</term><term>Chlorocebus aethiops</term><term>Neutralization Tests</term><term>Reproducibility of Results</term><term>SARS Virus (immunology)</term><term>Severe Acute Respiratory Syndrome (immunology)</term><term>Severe Acute Respiratory Syndrome (virology)</term><term>Vero Cells</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Anticorps antiviraux (analyse)</term><term>Anticorps antiviraux (immunologie)</term><term>Cellules Vero</term><term>Reproductibilité des résultats</term><term>Syndrome respiratoire aigu sévère (immunologie)</term><term>Syndrome respiratoire aigu sévère (virologie)</term><term>Tests de neutralisation</term><term>Virus du SRAS (immunologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Viral</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Anticorps antiviraux</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiviraux</term><term>Syndrome respiratoire aigu sévère</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>SARS Virus</term><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Syndrome respiratoire aigu sévère</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Severe Acute Respiratory Syndrome</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Chlorocebus aethiops</term><term>Neutralization Tests</term><term>Reproducibility of Results</term><term>Vero Cells</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Cellules Vero</term><term>Reproductibilité des résultats</term><term>Tests de neutralisation</term></keywords></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. 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></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Arita, I" uniqKey="Arita I">I. Arita</name></author><author><name sortKey="Kojima, K" uniqKey="Kojima K">K. Kojima</name></author><author><name sortKey="Nakane, M" uniqKey="Nakane M">M. Nakane</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Buchholz, U J" uniqKey="Buchholz U">U.J. Buchholz</name></author><author><name sortKey="Bukreyev, A" uniqKey="Bukreyev A">A. 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Leung</name></author></analytic></biblStruct></listBibl></div1></back></TEI><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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