Distinction of subtype-specific antibodies against European porcine influenza viruses by indirect ELISA based on recombinant hemagglutinin protein fragment-1
BACKGROUND: Serological investigations of swine influenza virus infections and epidemiological conclusions thereof are challenging due to the complex and regionally variable pattern of co-circulating viral subtypes and lineages and varying vaccination regimes. Detection of subtype-specific antibodie...
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| Veröffentlicht in: | Virology journal 2013-07, Vol.10 (1), p.246-246, Article 246 |
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| creator | Zhao, Na Lange, Elke Kubald, Sybille Grund, Christian Beer, Martin Harder, Timm C |
| description | BACKGROUND: Serological investigations of swine influenza virus infections and epidemiological conclusions thereof are challenging due to the complex and regionally variable pattern of co-circulating viral subtypes and lineages and varying vaccination regimes. Detection of subtype-specific antibodies currently depends on hemagglutination inhibition (HI) assays which are difficult to standardize and unsuitable for large scale investigations. METHODS: The nucleocapsid protein (NP) and HA1 fragments of the hemagglutinin protein (HA) of five different lineages (H1N1av, H1N1pdm, H1pdmN2, H1N2, H3N2) of swine influenza viruses were bacterially expressed and used as diagnostic antigens in indirect ELISA. RESULTS: Proteins were co-translationally mono-biotinylated and refolded in vitro into an antigenically authentic conformation. Western blotting and indirect ELISA revealed highly subtype-specific antigenic characteristics of the recombinant HA1 proteins although some cross reactivity especially among antigens of the H1 subtype were evident. Discrimination of antibodies directed against four swine influenza virus subtypes co-circulating in Germany was feasible using the indirect ELISA format. CONCLUSIONS: Bacterially expressed recombinant NP and HA1 swine influenza virus proteins served as antigens in indirect ELISAs and provided an alternative to commercial blocking NP ELISA and HI assays concerning generic (NP-specific) and HA subtype-specific sero-diagnostics, respectively, on a herd basis. |
| doi_str_mv | 10.1186/1743-422X-10-246 |
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Detection of subtype-specific antibodies currently depends on hemagglutination inhibition (HI) assays which are difficult to standardize and unsuitable for large scale investigations. METHODS: The nucleocapsid protein (NP) and HA1 fragments of the hemagglutinin protein (HA) of five different lineages (H1N1av, H1N1pdm, H1pdmN2, H1N2, H3N2) of swine influenza viruses were bacterially expressed and used as diagnostic antigens in indirect ELISA. RESULTS: Proteins were co-translationally mono-biotinylated and refolded in vitro into an antigenically authentic conformation. Western blotting and indirect ELISA revealed highly subtype-specific antigenic characteristics of the recombinant HA1 proteins although some cross reactivity especially among antigens of the H1 subtype were evident. Discrimination of antibodies directed against four swine influenza virus subtypes co-circulating in Germany was feasible using the indirect ELISA format. CONCLUSIONS: Bacterially expressed recombinant NP and HA1 swine influenza virus proteins served as antigens in indirect ELISAs and provided an alternative to commercial blocking NP ELISA and HI assays concerning generic (NP-specific) and HA subtype-specific sero-diagnostics, respectively, on a herd basis.</description><identifier>ISSN: 1743-422X</identifier><identifier>EISSN: 1743-422X</identifier><identifier>DOI: 10.1186/1743-422X-10-246</identifier><identifier>PMID: 23898799</identifier><language>eng</language><publisher>England: Springer-Verlag</publisher><subject>Animals ; Antibodies ; Antibodies, Viral - blood ; antigens ; Antigens, Viral - genetics ; cross reaction ; Diagnosis ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - methods ; Epidemiology ; Genetic aspects ; Germany ; hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus - genetics ; hemagglutinins ; herds ; Immunity ; Influenza A virus ; Influenza A virus - classification ; Influenza A virus - immunology ; Influenza A virus - isolation & purification ; nucleocapsid proteins ; Orthomyxoviridae Infections - immunology ; Orthomyxoviridae Infections - veterinary ; Orthomyxoviridae Infections - virology ; Physiological aspects ; Proteins ; Recombinant Proteins - genetics ; RNA-Binding Proteins - genetics ; Swine ; Swine Diseases - immunology ; Swine Diseases - virology ; Swine influenza ; swine influenza virus ; vaccination ; Viral antibodies ; Viral Core Proteins - genetics ; Viral proteins ; Western blotting</subject><ispartof>Virology journal, 2013-07, Vol.10 (1), p.246-246, Article 246</ispartof><rights>COPYRIGHT 2013 BioMed Central Ltd.</rights><rights>Copyright ©2013 Zhao et al.; licensee BioMed Central Ltd. 2013 Zhao et al.; licensee BioMed Central Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b647t-37cf6ba8aaaf41d0201e8bacd9c188473010b848774b250cd3e72296aca038d33</citedby><cites>FETCH-LOGICAL-b647t-37cf6ba8aaaf41d0201e8bacd9c188473010b848774b250cd3e72296aca038d33</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3733666/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3733666/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,725,778,782,862,883,27907,27908,53774,53776</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23898799$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhao, Na</creatorcontrib><creatorcontrib>Lange, Elke</creatorcontrib><creatorcontrib>Kubald, Sybille</creatorcontrib><creatorcontrib>Grund, Christian</creatorcontrib><creatorcontrib>Beer, Martin</creatorcontrib><creatorcontrib>Harder, Timm C</creatorcontrib><title>Distinction of subtype-specific antibodies against European porcine influenza viruses by indirect ELISA based on recombinant hemagglutinin protein fragment-1</title><title>Virology journal</title><addtitle>Virol J</addtitle><description>BACKGROUND: Serological investigations of swine influenza virus infections and epidemiological conclusions thereof are challenging due to the complex and regionally variable pattern of co-circulating viral subtypes and lineages and varying vaccination regimes. Detection of subtype-specific antibodies currently depends on hemagglutination inhibition (HI) assays which are difficult to standardize and unsuitable for large scale investigations. METHODS: The nucleocapsid protein (NP) and HA1 fragments of the hemagglutinin protein (HA) of five different lineages (H1N1av, H1N1pdm, H1pdmN2, H1N2, H3N2) of swine influenza viruses were bacterially expressed and used as diagnostic antigens in indirect ELISA. RESULTS: Proteins were co-translationally mono-biotinylated and refolded in vitro into an antigenically authentic conformation. Western blotting and indirect ELISA revealed highly subtype-specific antigenic characteristics of the recombinant HA1 proteins although some cross reactivity especially among antigens of the H1 subtype were evident. Discrimination of antibodies directed against four swine influenza virus subtypes co-circulating in Germany was feasible using the indirect ELISA format. CONCLUSIONS: Bacterially expressed recombinant NP and HA1 swine influenza virus proteins served as antigens in indirect ELISAs and provided an alternative to commercial blocking NP ELISA and HI assays concerning generic (NP-specific) and HA subtype-specific sero-diagnostics, respectively, on a herd basis.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Antibodies, Viral - blood</subject><subject>antigens</subject><subject>Antigens, Viral - genetics</subject><subject>cross reaction</subject><subject>Diagnosis</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Epidemiology</subject><subject>Genetic aspects</subject><subject>Germany</subject><subject>hemagglutination</subject><subject>Hemagglutinin Glycoproteins, Influenza Virus - genetics</subject><subject>hemagglutinins</subject><subject>herds</subject><subject>Immunity</subject><subject>Influenza A virus</subject><subject>Influenza A virus - classification</subject><subject>Influenza A virus - immunology</subject><subject>Influenza A virus - isolation & purification</subject><subject>nucleocapsid proteins</subject><subject>Orthomyxoviridae Infections - immunology</subject><subject>Orthomyxoviridae Infections - veterinary</subject><subject>Orthomyxoviridae Infections - virology</subject><subject>Physiological aspects</subject><subject>Proteins</subject><subject>Recombinant Proteins - genetics</subject><subject>RNA-Binding Proteins - genetics</subject><subject>Swine</subject><subject>Swine Diseases - immunology</subject><subject>Swine Diseases - virology</subject><subject>Swine influenza</subject><subject>swine influenza virus</subject><subject>vaccination</subject><subject>Viral antibodies</subject><subject>Viral Core Proteins - genetics</subject><subject>Viral proteins</subject><subject>Western blotting</subject><issn>1743-422X</issn><issn>1743-422X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkk2LFDEQhhtR3HX17kkbvOih13xNkr4Iw7jqwIDguOAtJOl0b6Q7mU26F8f_4n-1hlmHHVhh6UOaylNvVeqtoniJ0TnGkr_HgtGKEfKjwqgijD8qTg-hx3f-T4pnOf9EiBIu6qfFCaGylqKuT4s_H30efbCjj6GMbZknM243rsobZ33rbanD6E1svMul7rQPeSwvphQ3TodyE5P1wZU-tP3kwm9d3vg0ZUDNFoKNT84Cvlqu56XR2TUlFIFYHIwPIFxeuUF3XT9BBx7kUhwdnG3S3eDCWOHnxZNW99m9uD3PistPF98XX6rV18_LxXxVGc7EWFFhW2601Fq3DDeIIOyk0bapLZaSCYowMpJJIZghM2Qb6gQhNddWIyobSs-KD3vdzWQG11gonnSvNskPOm1V1F4d3wR_pbp4o6iglHMOAou9gPHxPwLHNzADtbNH7exRGClwD1Te3raR4vXk8qgGn63rex1cnDIkzCgjjPEHoIwItNuR2QNQLCmnjEtA3-zRTvdOga0RmrU7XM2hNK8Zkxio83so-Bo3eBuDaz3EjxLeHSUAM7pfY6ennNVy_e2YRXvWpphzcu1hiDCk3YPuG9uru-4dEv5tOQCv90Cro9Jd8lldrmFJZggRLAUsyF-MRghf</recordid><startdate>20130730</startdate><enddate>20130730</enddate><creator>Zhao, Na</creator><creator>Lange, Elke</creator><creator>Kubald, Sybille</creator><creator>Grund, Christian</creator><creator>Beer, Martin</creator><creator>Harder, Timm C</creator><general>Springer-Verlag</general><general>BioMed Central Ltd</general><general>BioMed Central</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>7X8</scope><scope>7U9</scope><scope>H94</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20130730</creationdate><title>Distinction of subtype-specific antibodies against European porcine influenza viruses by indirect ELISA based on recombinant hemagglutinin protein fragment-1</title><author>Zhao, Na ; Lange, Elke ; Kubald, Sybille ; Grund, Christian ; Beer, Martin ; Harder, Timm C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b647t-37cf6ba8aaaf41d0201e8bacd9c188473010b848774b250cd3e72296aca038d33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Antibodies, Viral - blood</topic><topic>antigens</topic><topic>Antigens, Viral - genetics</topic><topic>cross reaction</topic><topic>Diagnosis</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Epidemiology</topic><topic>Genetic aspects</topic><topic>Germany</topic><topic>hemagglutination</topic><topic>Hemagglutinin Glycoproteins, Influenza Virus - genetics</topic><topic>hemagglutinins</topic><topic>herds</topic><topic>Immunity</topic><topic>Influenza A virus</topic><topic>Influenza A virus - classification</topic><topic>Influenza A virus - immunology</topic><topic>Influenza A virus - isolation & purification</topic><topic>nucleocapsid proteins</topic><topic>Orthomyxoviridae Infections - immunology</topic><topic>Orthomyxoviridae Infections - veterinary</topic><topic>Orthomyxoviridae Infections - virology</topic><topic>Physiological aspects</topic><topic>Proteins</topic><topic>Recombinant Proteins - genetics</topic><topic>RNA-Binding Proteins - genetics</topic><topic>Swine</topic><topic>Swine Diseases - immunology</topic><topic>Swine Diseases - virology</topic><topic>Swine influenza</topic><topic>swine influenza virus</topic><topic>vaccination</topic><topic>Viral antibodies</topic><topic>Viral Core Proteins - genetics</topic><topic>Viral proteins</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhao, Na</creatorcontrib><creatorcontrib>Lange, Elke</creatorcontrib><creatorcontrib>Kubald, Sybille</creatorcontrib><creatorcontrib>Grund, Christian</creatorcontrib><creatorcontrib>Beer, Martin</creatorcontrib><creatorcontrib>Harder, Timm C</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>MEDLINE - Academic</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Virology journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhao, Na</au><au>Lange, Elke</au><au>Kubald, Sybille</au><au>Grund, Christian</au><au>Beer, Martin</au><au>Harder, Timm C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Distinction of subtype-specific antibodies against European porcine influenza viruses by indirect ELISA based on recombinant hemagglutinin protein fragment-1</atitle><jtitle>Virology journal</jtitle><addtitle>Virol J</addtitle><date>2013-07-30</date><risdate>2013</risdate><volume>10</volume><issue>1</issue><spage>246</spage><epage>246</epage><pages>246-246</pages><artnum>246</artnum><issn>1743-422X</issn><eissn>1743-422X</eissn><abstract>BACKGROUND: Serological investigations of swine influenza virus infections and epidemiological conclusions thereof are challenging due to the complex and regionally variable pattern of co-circulating viral subtypes and lineages and varying vaccination regimes. Detection of subtype-specific antibodies currently depends on hemagglutination inhibition (HI) assays which are difficult to standardize and unsuitable for large scale investigations. METHODS: The nucleocapsid protein (NP) and HA1 fragments of the hemagglutinin protein (HA) of five different lineages (H1N1av, H1N1pdm, H1pdmN2, H1N2, H3N2) of swine influenza viruses were bacterially expressed and used as diagnostic antigens in indirect ELISA. RESULTS: Proteins were co-translationally mono-biotinylated and refolded in vitro into an antigenically authentic conformation. Western blotting and indirect ELISA revealed highly subtype-specific antigenic characteristics of the recombinant HA1 proteins although some cross reactivity especially among antigens of the H1 subtype were evident. Discrimination of antibodies directed against four swine influenza virus subtypes co-circulating in Germany was feasible using the indirect ELISA format. CONCLUSIONS: Bacterially expressed recombinant NP and HA1 swine influenza virus proteins served as antigens in indirect ELISAs and provided an alternative to commercial blocking NP ELISA and HI assays concerning generic (NP-specific) and HA subtype-specific sero-diagnostics, respectively, on a herd basis.</abstract><cop>England</cop><pub>Springer-Verlag</pub><pmid>23898799</pmid><doi>10.1186/1743-422X-10-246</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central Open Access; Springer Nature OA Free Journals; PubMed Central; SpringerLink Journals - AutoHoldings |
| subjects | Animals Antibodies Antibodies, Viral - blood antigens Antigens, Viral - genetics cross reaction Diagnosis Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods Epidemiology Genetic aspects Germany hemagglutination Hemagglutinin Glycoproteins, Influenza Virus - genetics hemagglutinins herds Immunity Influenza A virus Influenza A virus - classification Influenza A virus - immunology Influenza A virus - isolation & purification nucleocapsid proteins Orthomyxoviridae Infections - immunology Orthomyxoviridae Infections - veterinary Orthomyxoviridae Infections - virology Physiological aspects Proteins Recombinant Proteins - genetics RNA-Binding Proteins - genetics Swine Swine Diseases - immunology Swine Diseases - virology Swine influenza swine influenza virus vaccination Viral antibodies Viral Core Proteins - genetics Viral proteins Western blotting |
| title | Distinction of subtype-specific antibodies against European porcine influenza viruses by indirect ELISA based on recombinant hemagglutinin protein fragment-1 |
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