Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe...
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| Veröffentlicht in: | Virology journal 2015-08, Vol.12 (1), p.117-117, Article 117 |
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| creator | Yu, Fuxun Du, Yanhua Huang, Xueyong Ma, Hong Xu, Bianli Adungo, Ferdinard Hayasaka, Daisuke Buerano, Corazon C Morita, Kouichi |
| description | Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe and affordable serodiagnostic methods for proper clinical management of infected patients.
The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system.
The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively.
The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations. |
| doi_str_mv | 10.1186/s12985-015-0350-0 |
| format | Article |
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The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system.
The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively.
The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations.</description><identifier>ISSN: 1743-422X</identifier><identifier>EISSN: 1743-422X</identifier><identifier>DOI: 10.1186/s12985-015-0350-0</identifier><identifier>PMID: 26239826</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Diagnosis ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - methods ; Enzymes ; Epidemiology ; Escherichia coli ; Gene Expression ; Genetic aspects ; Genetic engineering ; Health aspects ; Humans ; Immunoglobulin G ; Immunoglobulin G - blood ; Immunoglobulin G - immunology ; Immunoglobulin M - blood ; Immunoglobulin M - immunology ; Methodology ; Nucleocapsid Proteins - genetics ; Nucleocapsid Proteins - immunology ; Nucleocapsid Proteins - isolation & purification ; Phlebotomus Fever - diagnosis ; Phlebotomus Fever - immunology ; Phlebotomus Fever - virology ; Phlebovirus - genetics ; Phlebovirus - immunology ; Recombinant Proteins ; Thrombocytopenia ; Virus diseases</subject><ispartof>Virology journal, 2015-08, Vol.12 (1), p.117-117, Article 117</ispartof><rights>COPYRIGHT 2015 BioMed Central Ltd.</rights><rights>Copyright BioMed Central 2015</rights><rights>Yu et al. 2015</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c638t-e4030b4f928f0bb2c5033c00403b0765001d34d90fecd6ead233f67dbe78762f3</citedby><cites>FETCH-LOGICAL-c638t-e4030b4f928f0bb2c5033c00403b0765001d34d90fecd6ead233f67dbe78762f3</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/PMC4524020/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4524020/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27923,27924,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26239826$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yu, Fuxun</creatorcontrib><creatorcontrib>Du, Yanhua</creatorcontrib><creatorcontrib>Huang, Xueyong</creatorcontrib><creatorcontrib>Ma, Hong</creatorcontrib><creatorcontrib>Xu, Bianli</creatorcontrib><creatorcontrib>Adungo, Ferdinard</creatorcontrib><creatorcontrib>Hayasaka, Daisuke</creatorcontrib><creatorcontrib>Buerano, Corazon C</creatorcontrib><creatorcontrib>Morita, Kouichi</creatorcontrib><title>Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA</title><title>Virology journal</title><addtitle>Virol J</addtitle><description>Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe and affordable serodiagnostic methods for proper clinical management of infected patients.
The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system.
The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively.
The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations.</description><subject>Diagnosis</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Enzymes</subject><subject>Epidemiology</subject><subject>Escherichia coli</subject><subject>Gene Expression</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Health aspects</subject><subject>Humans</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulin G - blood</subject><subject>Immunoglobulin G - immunology</subject><subject>Immunoglobulin M - blood</subject><subject>Immunoglobulin M - immunology</subject><subject>Methodology</subject><subject>Nucleocapsid Proteins - genetics</subject><subject>Nucleocapsid Proteins - immunology</subject><subject>Nucleocapsid Proteins - isolation & purification</subject><subject>Phlebotomus Fever - diagnosis</subject><subject>Phlebotomus Fever - immunology</subject><subject>Phlebotomus Fever - virology</subject><subject>Phlebovirus - genetics</subject><subject>Phlebovirus - immunology</subject><subject>Recombinant Proteins</subject><subject>Thrombocytopenia</subject><subject>Virus diseases</subject><issn>1743-422X</issn><issn>1743-422X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNptkl1rFDEUhgdR7If-AG8k4I1eTM3HfN4IS2nrworgVvEuZJKT3ZSZZJpkVvd_9QeaZdvaFQnhhJPnPSc5vFn2huAzQprqYyC0bcock7RZiXP8LDsmdcHygtKfz5-cj7KTEG4wZrSq25fZEa0oaxtaHWd3s3HsjRTROIucRh6kGzpjhY0owAY8IL0L6JeJaxTXPt06uY1uBGsEClurUgrQxvgpIDvJHpwUYzAKjd5FMBZp55MQkIII8qHP8vJ6-SMPI0ijjUTraRAWzVdXSFiV4pcUo-mcMhBQt0XGKpOeFtHFYr6cvcpeaNEHeH0fT7PvlxfX55_zxder-flskcuKNTGHAjPcFbqljcZdR2WJGZMYp3SH66rEmChWqBZrkKoCoShjuqpVB3VTV1Sz0-zTvu44dQMoCTZ60fPRm0H4LXfC8MMba9Z85Ta8KGmBKU4F3t8X8O52ghD5YIKEvhcW3BQ4qRPVtGVDE_ruH_TGTd6m7yWqwVVLcUn-UivRAzdWu9RX7oryWVmQkjaUFok6-w-VloLBSGdBm5Q_EHw4ECQmwu-4ElMIfL78dsiSPSu9C8GDfpwHwXxnS763JU-25Dtb8t0c3j4d5KPiwYfsD0N03yE</recordid><startdate>20150804</startdate><enddate>20150804</enddate><creator>Yu, Fuxun</creator><creator>Du, Yanhua</creator><creator>Huang, Xueyong</creator><creator>Ma, Hong</creator><creator>Xu, Bianli</creator><creator>Adungo, Ferdinard</creator><creator>Hayasaka, Daisuke</creator><creator>Buerano, Corazon C</creator><creator>Morita, Kouichi</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><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>3V.</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>COVID</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20150804</creationdate><title>Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA</title><author>Yu, Fuxun ; Du, Yanhua ; Huang, Xueyong ; Ma, Hong ; Xu, Bianli ; Adungo, Ferdinard ; Hayasaka, Daisuke ; Buerano, Corazon C ; Morita, Kouichi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c638t-e4030b4f928f0bb2c5033c00403b0765001d34d90fecd6ead233f67dbe78762f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Diagnosis</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Enzymes</topic><topic>Epidemiology</topic><topic>Escherichia coli</topic><topic>Gene Expression</topic><topic>Genetic aspects</topic><topic>Genetic engineering</topic><topic>Health aspects</topic><topic>Humans</topic><topic>Immunoglobulin G</topic><topic>Immunoglobulin G - blood</topic><topic>Immunoglobulin G - immunology</topic><topic>Immunoglobulin M - blood</topic><topic>Immunoglobulin M - immunology</topic><topic>Methodology</topic><topic>Nucleocapsid Proteins - genetics</topic><topic>Nucleocapsid Proteins - immunology</topic><topic>Nucleocapsid Proteins - isolation & purification</topic><topic>Phlebotomus Fever - diagnosis</topic><topic>Phlebotomus Fever - immunology</topic><topic>Phlebotomus Fever - virology</topic><topic>Phlebovirus - genetics</topic><topic>Phlebovirus - immunology</topic><topic>Recombinant Proteins</topic><topic>Thrombocytopenia</topic><topic>Virus diseases</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yu, Fuxun</creatorcontrib><creatorcontrib>Du, Yanhua</creatorcontrib><creatorcontrib>Huang, Xueyong</creatorcontrib><creatorcontrib>Ma, Hong</creatorcontrib><creatorcontrib>Xu, Bianli</creatorcontrib><creatorcontrib>Adungo, Ferdinard</creatorcontrib><creatorcontrib>Hayasaka, Daisuke</creatorcontrib><creatorcontrib>Buerano, Corazon C</creatorcontrib><creatorcontrib>Morita, Kouichi</creatorcontrib><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>ProQuest Central (Corporate)</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Coronavirus Research Database</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - 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>Yu, Fuxun</au><au>Du, Yanhua</au><au>Huang, Xueyong</au><au>Ma, Hong</au><au>Xu, Bianli</au><au>Adungo, Ferdinard</au><au>Hayasaka, Daisuke</au><au>Buerano, Corazon C</au><au>Morita, Kouichi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA</atitle><jtitle>Virology journal</jtitle><addtitle>Virol J</addtitle><date>2015-08-04</date><risdate>2015</risdate><volume>12</volume><issue>1</issue><spage>117</spage><epage>117</epage><pages>117-117</pages><artnum>117</artnum><issn>1743-422X</issn><eissn>1743-422X</eissn><abstract>Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease that was first reported in China in 2011. It is caused by SFTS virus (SFTSV) which is a member of the Phlebovirus genus in the Bunyaviridae family. SFTSV has been classified as a BSL3 pathogen. There is a need to develop safe and affordable serodiagnostic methods for proper clinical management of infected patients.
The full length nucleocapsid (N) gene of SFTSV Yamaguchi strain was amplified by RT-PCR and cloned to an expression vector pQE30. The recombinant (r) SFTSV-N protein was expressed by using Escherichia coli (E. coli) expression system and purified under native conditions. rSFTSV-N protein based indirect IgG and IgM enzyme linked immunosorbent assay (ELISA) systems were established to detect specific human IgG and IgM antibodies, respectively. One hundred fifteen serum samples from clinically suspected-SFTS patients were used to evaluate the newly established systems and the results were compared with the total antibody detecting sandwich ELISA system.
The native form of recombinant (r) SFTSV-N protein was expressed and purified. Application of the rSFTSV-N protein based indirect IgG ELISA to the 115 serum samples showed results that perfectly matched those of the total antibody sandwich ELISA with a sensitivity and specificity of 100 %. The rSFTSV-N protein based indirect IgM ELISA missed 8 positive samples that were detected by the total antibody sandwich ELISA. The sensitivity and specificity of rSFTSV-N-IgM capture ELISA were 90.59 and 100 %, respectively.
The rSFTSV-N protein is highly immunoreactive and a good target for use as an assay antigen in laboratory diagnosis. Its preparation is simpler in comparison with that used for the total antibody sandwich system. Our rSFTSV-N protein-based IgG and IgM ELISA systems have the advantage of distinguishing two types of antibodies and require small volume of serum sample only. They are safe to use for diagnosis of SFTS virus infection and especially fit in large-scale epidemiological investigations.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>26239826</pmid><doi>10.1186/s12985-015-0350-0</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; DOAJ Directory of Open Access Journals; PubMed Central Open Access; Springer Nature OA Free Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; SpringerLink Journals - AutoHoldings |
| subjects | Diagnosis Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods Enzymes Epidemiology Escherichia coli Gene Expression Genetic aspects Genetic engineering Health aspects Humans Immunoglobulin G Immunoglobulin G - blood Immunoglobulin G - immunology Immunoglobulin M - blood Immunoglobulin M - immunology Methodology Nucleocapsid Proteins - genetics Nucleocapsid Proteins - immunology Nucleocapsid Proteins - isolation & purification Phlebotomus Fever - diagnosis Phlebotomus Fever - immunology Phlebotomus Fever - virology Phlebovirus - genetics Phlebovirus - immunology Recombinant Proteins Thrombocytopenia Virus diseases |
| title | Application of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for the detection of SFTSV-specific human IgG and IgM antibodies by indirect ELISA |
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