A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein
Identifieur interne : 000B94 ( Istex/Corpus ); précédent : 000B93; suivant : 000B95A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein
Auteurs : L. Gerentes ; N. Kessler ; M. AymardSource :
- Journal of Virological Methods [ 0166-0934 ] ; 1998.
English descriptors
- Teeft :
- Accurate quantitation, Adsorption, Antibody response, Antigen detection, Antigenic, Antigenic reactivity, Blank controls, Blank reaction, Chicken erythrocytes, Current vaccines, Detector antibodies, Detector antibody, Elisa, Elisa immunocapture assay, Elsevier science, Enzymatic activity, European pharmacopoeia, Gerentes, Hemagglutinin, Immunocapture, Immunocapture checkerboard titration, Inhibition test, Inhibition tests, Johansson, Kilbourne, Linear part, Microtiter plates, Monoclonal antibodies, Monoclonal antibody, Neuraminidase, Optimal dilution, Optimal dilutiond, Page integration, Pasteur merieux connaught, Polyclonal, Polyclonal rabbit, Precise estimation, Protein quantitation, Prototype strains, Quantitation, Rabbit antiserum, Rapid diagnosis, Reference antigen, Reference curve, Room temperature, Sample dilutions, Saturation step, Standard curve, Subunit vaccines, Trivalent, Trivalent suspension, Trivalent vaccine doses, Trivalent vaccines, Tween ether treatment, Vaccine, Validation, Validation method, Viral, Viral samples, Virol, Virological, Virological methods, Virus, Virus hemagglutinin, Virus infection, Virus neuraminidase, Whole virus.
Abstract
Abstract: Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.
Url:
DOI: 10.1016/S0166-0934(98)00056-1
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ISTEX:FA9386160BBC83FC8E17C1A5ED26B79A7A0D5C01Le document en format XML
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Gerentes</name><affiliation><mods:affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</mods:affiliation></affiliation></author><author><name sortKey="Kessler, N" sort="Kessler, N" uniqKey="Kessler N" first="N" last="Kessler">N. Kessler</name><affiliation><mods:affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</mods:affiliation></affiliation></author><author><name sortKey="Aymard, M" sort="Aymard, M" uniqKey="Aymard M" first="M" last="Aymard">M. Aymard</name><affiliation><mods:affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998">1998</date><biblScope unit="volume">73</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="185">185</biblScope><biblScope unit="page" to="195">195</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Accurate quantitation</term><term>Adsorption</term><term>Antibody response</term><term>Antigen detection</term><term>Antigenic</term><term>Antigenic reactivity</term><term>Blank controls</term><term>Blank reaction</term><term>Chicken erythrocytes</term><term>Current vaccines</term><term>Detector antibodies</term><term>Detector antibody</term><term>Elisa</term><term>Elisa immunocapture assay</term><term>Elsevier science</term><term>Enzymatic activity</term><term>European pharmacopoeia</term><term>Gerentes</term><term>Hemagglutinin</term><term>Immunocapture</term><term>Immunocapture checkerboard titration</term><term>Inhibition test</term><term>Inhibition tests</term><term>Johansson</term><term>Kilbourne</term><term>Linear part</term><term>Microtiter plates</term><term>Monoclonal antibodies</term><term>Monoclonal antibody</term><term>Neuraminidase</term><term>Optimal dilution</term><term>Optimal dilutiond</term><term>Page integration</term><term>Pasteur merieux connaught</term><term>Polyclonal</term><term>Polyclonal rabbit</term><term>Precise estimation</term><term>Protein quantitation</term><term>Prototype strains</term><term>Quantitation</term><term>Rabbit antiserum</term><term>Rapid diagnosis</term><term>Reference antigen</term><term>Reference curve</term><term>Room temperature</term><term>Sample dilutions</term><term>Saturation step</term><term>Standard curve</term><term>Subunit vaccines</term><term>Trivalent</term><term>Trivalent suspension</term><term>Trivalent vaccine doses</term><term>Trivalent vaccines</term><term>Tween ether treatment</term><term>Vaccine</term><term>Validation</term><term>Validation method</term><term>Viral</term><term>Viral samples</term><term>Virol</term><term>Virological</term><term>Virological methods</term><term>Virus</term><term>Virus hemagglutinin</term><term>Virus infection</term><term>Virus neuraminidase</term><term>Whole virus</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>quantitation</json:string><json:string>neuraminidase</json:string><json:string>vaccine</json:string><json:string>elisa</json:string><json:string>gerentes</json:string><json:string>polyclonal</json:string><json:string>kilbourne</json:string><json:string>antigenic</json:string><json:string>trivalent</json:string><json:string>virological</json:string><json:string>hemagglutinin</json:string><json:string>virological methods</json:string><json:string>johansson</json:string><json:string>immunocapture</json:string><json:string>virol</json:string><json:string>adsorption</json:string><json:string>viral</json:string><json:string>linear part</json:string><json:string>whole virus</json:string><json:string>reference curve</json:string><json:string>validation</json:string><json:string>page integration</json:string><json:string>antigen detection</json:string><json:string>enzymatic activity</json:string><json:string>inhibition test</json:string><json:string>validation method</json:string><json:string>elisa immunocapture assay</json:string><json:string>antigenic reactivity</json:string><json:string>blank controls</json:string><json:string>chicken erythrocytes</json:string><json:string>reference antigen</json:string><json:string>sample dilutions</json:string><json:string>virus neuraminidase</json:string><json:string>polyclonal rabbit</json:string><json:string>rabbit antiserum</json:string><json:string>room temperature</json:string><json:string>optimal dilutiond</json:string><json:string>precise estimation</json:string><json:string>inhibition tests</json:string><json:string>microtiter plates</json:string><json:string>optimal dilution</json:string><json:string>immunocapture checkerboard titration</json:string><json:string>european pharmacopoeia</json:string><json:string>blank reaction</json:string><json:string>tween ether treatment</json:string><json:string>elsevier science</json:string><json:string>detector antibody</json:string><json:string>accurate quantitation</json:string><json:string>current vaccines</json:string><json:string>trivalent vaccine doses</json:string><json:string>saturation step</json:string><json:string>prototype strains</json:string><json:string>detector antibodies</json:string><json:string>standard curve</json:string><json:string>monoclonal antibody</json:string><json:string>viral samples</json:string><json:string>pasteur merieux connaught</json:string><json:string>trivalent vaccines</json:string><json:string>trivalent suspension</json:string><json:string>virus infection</json:string><json:string>subunit vaccines</json:string><json:string>protein quantitation</json:string><json:string>rapid diagnosis</json:string><json:string>antibody response</json:string><json:string>virus hemagglutinin</json:string><json:string>monoclonal antibodies</json:string><json:string>virus</json:string></teeft></keywords><author><json:item><name>L Gerentes</name><affiliations><json:string>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</json:string></affiliations></json:item><json:item><name>N Kessler</name><affiliations><json:string>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</json:string></affiliations></json:item><json:item><name>M Aymard</name><affiliations><json:string>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Influenza virus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Neuraminidase quantitation</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>ELISA immunocapture assay</value></json:item></subject><arkIstex>ark:/67375/6H6-V6JM3DXF-4</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.</abstract><qualityIndicators><score>7.861</score><pdfWordCount>4217</pdfWordCount><pdfCharCount>28096</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>11</pdfPageCount><pdfPageSize>536 x 674 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>137</abstractWordCount><abstractCharCount>902</abstractCharCount><keywordCount>3</keywordCount></qualityIndicators><title>A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</title><pmid><json:string>9766889</json:string></pmid><pii><json:string>S0166-0934(98)00056-1</json:string></pii><genre><json:string>research-article</json:string></genre><host><title>Journal of Virological Methods</title><language><json:string>unknown</json:string></language><publicationDate>1998</publicationDate><issn><json:string>0166-0934</json:string></issn><pii><json:string>S0166-0934(00)X0041-9</json:string></pii><volume>73</volume><issue>2</issue><pages><first>185</first><last>195</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1998</json:string><json:string>1980s</json:string></date><geogName></geogName><orgName><json:string>France Received</json:string><json:string>Merieux Foun´</json:string></orgName><orgName_funder><json:string>Merieux Foun´</json:string></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Chalumeau</json:string><json:string>N. Kessler</json:string></persName><placeName><json:string>Singapore</json:string><json:string>Beijing</json:string><json:string>Mannheim</json:string><json:string>Harbin</json:string><json:string>Puri</json:string><json:string>France</json:string><json:string>Zagreb</json:string><json:string>Panama</json:string><json:string>Lyon</json:string><json:string>Johannesburg</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Couch et al., 1997</json:string><json:string>Kilbourne, 1969</json:string><json:string>Kida et al. (1982)</json:string><json:string>Webster et al., 1988</json:string><json:string>Schulmann et al., 1968</json:string><json:string>L. Gerentes et al.</json:string><json:string>Johansson and Kilbourne, 1993</json:string><json:string>Laver and Valentine, 1969</json:string><json:string>Kilbourne et al., 1968</json:string><json:string>Aymard-Henry et al. (1973)</json:string><json:string>Gerentes et al.</json:string><json:string>Palmer et al. (1975)</json:string><json:string>Johansson et al., 1995</json:string><json:string>Tka` cova and Vareckova, 1996</json:string><json:string>Johansson et al., 1989</json:string><json:string>Deroo et al., 1996</json:string><json:string>Kilbourne et al., 1995</json:string><json:string>Chomel et al., 1989</json:string><json:string>Schild et al., 1975</json:string><json:string>Coonrod et al., 1984</json:string><json:string>Aymard-Henry et al., 1973</json:string><json:string>Aymard et al., 1996</json:string><json:string>Wood et al., 1977</json:string><json:string>Laemmli, 1970</json:string><json:string>Hocart et al., 1995</json:string><json:string>Gerentes et al., 1996</json:string><json:string>Norrby, 1962</json:string><json:string>Lowry et al. (1951)</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-V6JM3DXF-4</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - virology</json:string><json:string>2 - biotechnology & applied microbiology</json:string><json:string>2 - biochemical research methods</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - virology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Immunology and Microbiology</json:string><json:string>3 - Virology</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences biologiques fondamentales et appliquees. psychologie</json:string><json:string>4 - psychologie. psychophysiologie</json:string></inist></categories><publicationDate>1998</publicationDate><copyrightDate>1998</copyrightDate><doi><json:string>10.1016/S0166-0934(98)00056-1</json:string></doi><id>FA9386160BBC83FC8E17C1A5ED26B79A7A0D5C01</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-V6JM3DXF-4/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-V6JM3DXF-4/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-V6JM3DXF-4/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a">A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>©1998 Elsevier Science B.V.</p></licence><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</p></availability><date>1998</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Fig. 1: NA/N2 reference curve in NA-EIA.</note><note type="content">Fig. 2: NA/N2 curve of trivalent dose (home made) in NA EIA.</note><note type="content">Table 1: Characterization of anti-neuraminidase MAbs generated against A/Beijing/32/92 H3N2</note><note type="content">Table 2: Characterization of specific polyclonal rabbit anti-sera raised against either purified whole X117 virus or purified X117 NA</note><note type="content">Table 3: Quality control of the saturation step in NA-EIA</note><note type="content">Table 4: Comparative suitability of polyclonal anti-X117 rabbit antiserum and polyclonal anti-purified NA rabbit antiserum as detector antibody in NA-EIA</note><note type="content">Table 5: Estimation of ST9D2 cross-reactivity in ELISA and NI tests with several A and B virus types and subtypes</note><note type="content">Table 6: Determination of NA-EIA precision</note><note type="content">Table 7: Determination of NA-EIA accuracy</note><note type="content">Table 8: Antigenic ELISA cross reactivity of ST9D2 MAb with H3N2 viruses</note><note type="content">Table 9: Evaluation of NA concentration in manufactured vaccines using NA-EIA</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</title><author xml:id="author-0000"><persName><forename type="first">L</forename><surname>Gerentes</surname></persName><note type="correspondence"><p>Corresponding author.</p></note><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation></author><author xml:id="author-0001"><persName><forename type="first">N</forename><surname>Kessler</surname></persName><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation></author><author xml:id="author-0002"><persName><forename type="first">M</forename><surname>Aymard</surname></persName><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation></author><idno type="istex">FA9386160BBC83FC8E17C1A5ED26B79A7A0D5C01</idno><idno type="ark">ark:/67375/6H6-V6JM3DXF-4</idno><idno type="DOI">10.1016/S0166-0934(98)00056-1</idno><idno type="PII">S0166-0934(98)00056-1</idno></analytic><monogr><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="pISSN">0166-0934</idno><idno type="PII">S0166-0934(00)X0041-9</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1998"></date><biblScope unit="volume">73</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="185">185</biblScope><biblScope unit="page" to="195">195</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1998</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.</p></abstract><textClass><keywords scheme="keyword"><list><head>Keywords</head><item><term>Influenza virus</term></item><item><term>Neuraminidase quantitation</term></item><item><term>ELISA immunocapture assay</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="1998-03-25">Modified</change><change when="1998">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-V6JM3DXF-4/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"><istex:entity SYSTEM="gr1" NDATA="IMAGE" name="gr1"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla"><item-info><jid>VIRMET</jid><aid>2445</aid><ce:pii>S0166-0934(98)00056-1</ce:pii><ce:doi>10.1016/S0166-0934(98)00056-1</ce:doi><ce:copyright year="1998" type="full-transfer">Elsevier Science B.V.</ce:copyright></item-info><head><ce:title>A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</ce:title><ce:author-group><ce:author><ce:given-name>L</ce:given-name><ce:surname>Gerentes</ce:surname><ce:cross-ref refid="CORR1">*</ce:cross-ref></ce:author><ce:author><ce:given-name>N</ce:given-name><ce:surname>Kessler</ce:surname></ce:author><ce:author><ce:given-name>M</ce:given-name><ce:surname>Aymard</ce:surname></ce:author><ce:affiliation><ce:textfn>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</ce:textfn></ce:affiliation><ce:correspondence id="CORR1"><ce:label>*</ce:label><ce:text>Corresponding author.</ce:text></ce:correspondence></ce:author-group><ce:date-received day="20" month="11" year="1997"></ce:date-received><ce:date-revised day="25" month="3" year="1998"></ce:date-revised><ce:date-accepted day="25" month="3" year="1998"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Influenza virus</ce:text></ce:keyword><ce:keyword><ce:text>Neuraminidase quantitation</ce:text></ce:keyword><ce:keyword><ce:text>ELISA immunocapture assay</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>A sensitive and specific ELISA immunocapture assay for rapid quantitation of influenza A/H3N2 neuraminidase protein</title></titleInfo><name type="personal"><namePart type="given">L</namePart><namePart type="family">Gerentes</namePart><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation><description>Corresponding author.</description><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">N</namePart><namePart type="family">Kessler</namePart><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">M</namePart><namePart type="family">Aymard</namePart><affiliation>W.H.O., National Influenza Centre, Laboratoire de Virologie, 8, Avenue Rockefeller, 69373, Lyon Cedex 08, France</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1998</dateIssued><dateModified encoding="w3cdtf">1998-03-25</dateModified><copyrightDate encoding="w3cdtf">1998</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: Both HA and NA proteins elicit antibodies which have been shown to be capable of altering the course of infection. Nevertheless, while influenza virus vaccine standardization involves hemagglutinin (HA) and neuraminidase (NA) in terms of antigenic characterization, only HA protein quantitation is undertaken. An immunocapture ELISA (EIA) is described for N2 NA quantitation, based on the use of a highly specific monoclonal antibody (MAb) for capturing NA and an anti-NA antiserum for antigen detection. The amounts of NA in samples were deduced from the standard curve established by using purified NA. The NA-EIA is specific and detects as a little as 7 ng/ml. The capture and detector antibodies directed against A/Beijing/32/92 NA were shown to react with H3N2 prototype strains used in current influenza vaccines, provided that an antigenically matched reference NA is used as standard.</abstract><note type="content">Fig. 1: NA/N2 reference curve in NA-EIA.</note><note type="content">Fig. 2: NA/N2 curve of trivalent dose (home made) in NA EIA.</note><note type="content">Table 1: Characterization of anti-neuraminidase MAbs generated against A/Beijing/32/92 H3N2</note><note type="content">Table 2: Characterization of specific polyclonal rabbit anti-sera raised against either purified whole X117 virus or purified X117 NA</note><note type="content">Table 3: Quality control of the saturation step in NA-EIA</note><note type="content">Table 4: Comparative suitability of polyclonal anti-X117 rabbit antiserum and polyclonal anti-purified NA rabbit antiserum as detector antibody in NA-EIA</note><note type="content">Table 5: Estimation of ST9D2 cross-reactivity in ELISA and NI tests with several A and B virus types and subtypes</note><note type="content">Table 6: Determination of NA-EIA precision</note><note type="content">Table 7: Determination of NA-EIA accuracy</note><note type="content">Table 8: Antigenic ELISA cross reactivity of ST9D2 MAb with H3N2 viruses</note><note type="content">Table 9: Evaluation of NA concentration in manufactured vaccines using NA-EIA</note><subject><genre>Keywords</genre><topic>Influenza virus</topic><topic>Neuraminidase quantitation</topic><topic>ELISA immunocapture assay</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Virological Methods</title></titleInfo><titleInfo type="abbreviated"><title>VIRMET</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1998</dateIssued></originInfo><identifier type="ISSN">0166-0934</identifier><identifier type="PII">S0166-0934(00)X0041-9</identifier><part><date>1998</date><detail type="volume"><number>73</number><caption>vol.</caption></detail><detail type="issue"><number>2</number><caption>no.</caption></detail><extent unit="issue-pages"><start>123</start><end>246</end></extent><extent unit="pages"><start>185</start><end>195</end></extent></part></relatedItem><identifier type="istex">FA9386160BBC83FC8E17C1A5ED26B79A7A0D5C01</identifier><identifier type="ark">ark:/67375/6H6-V6JM3DXF-4</identifier><identifier type="DOI">10.1016/S0166-0934(98)00056-1</identifier><identifier type="PII">S0166-0934(98)00056-1</identifier><accessCondition type="use and reproduction" contentType="copyright">©1998 Elsevier Science B.V.</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource><recordOrigin>Elsevier Science B.V., ©1998</recordOrigin></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-V6JM3DXF-4/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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