Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection
Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to...
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description | Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. Here, we establish a reliable 1F5 based-icELISA that constitutes a promising diagnostic tool for control strategies and the prevention of ZIKV propagation. |
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There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. Here, we establish a reliable 1F5 based-icELISA that constitutes a promising diagnostic tool for control strategies and the prevention of ZIKV propagation.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0256220</identifier><identifier>PMID: 34403457</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Animals ; Antibodies, Monoclonal - biosynthesis ; Antibodies, Monoclonal - chemistry ; Antibodies, Monoclonal - isolation & purification ; Antibodies, Viral - biosynthesis ; Antibodies, Viral - chemistry ; Antibodies, Viral - isolation & purification ; Antigens, Viral - administration & dosage ; Antigens, Viral - genetics ; Antigens, Viral - immunology ; Binding, Competitive ; Biology and Life Sciences ; Chromatography ; Cloning, Molecular ; Congenital defects ; Cross-reaction ; Dengue fever ; Early Diagnosis ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - methods ; Enzyme-Linked Immunosorbent Assay - standards ; Enzymes ; Epitopes ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Flaviviridae ; Gene Expression ; Global health ; Guillain-Barre syndrome ; Health risks ; HEK293 Cells ; Humans ; Hybridomas - chemistry ; Hybridomas - immunology ; Immunization ; Infections ; Male ; Markers ; Medicine and Health Sciences ; Mice ; Mice, Inbred BALB C ; Monoclonal antibodies ; NS1 protein ; Protein Multimerization ; Proteins ; Public health ; Recombinant Proteins - administration & dosage ; Recombinant Proteins - genetics ; Recombinant Proteins - immunology ; Reproducibility of Results ; Research and Analysis Methods ; Sensitivity and Specificity ; Vaccines ; Vector-borne diseases ; Viral Nonstructural Proteins - administration & dosage ; Viral Nonstructural Proteins - genetics ; Viral Nonstructural Proteins - immunology ; Viruses ; West Nile virus ; Zika virus ; Zika Virus - genetics ; Zika Virus - immunology ; Zika Virus Infection - diagnosis ; Zika Virus Infection - immunology ; Zika Virus Infection - virology</subject><ispartof>PloS one, 2021-08, Vol.16 (8), p.e0256220-e0256220</ispartof><rights>COPYRIGHT 2021 Public Library of Science</rights><rights>2021 Roldán et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 Roldán et al 2021 Roldán et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c758t-f91b6680757d57d80ccf0bf4c3ca4a7cdfc38c949cb7b214767c123d5ef3d7223</citedby><cites>FETCH-LOGICAL-c758t-f91b6680757d57d80ccf0bf4c3ca4a7cdfc38c949cb7b214767c123d5ef3d7223</cites><orcidid>0000-0002-8307-2100</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8370630/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8370630/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34403457$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Ito, Etsuro</contributor><creatorcontrib>Roldán, Julieta S</creatorcontrib><creatorcontrib>Cassola, Alejandro</creatorcontrib><creatorcontrib>Castillo, Daniela S</creatorcontrib><title>Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Roldán, Julieta S</au><au>Cassola, Alejandro</au><au>Castillo, Daniela S</au><au>Ito, Etsuro</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2021-08-17</date><risdate>2021</risdate><volume>16</volume><issue>8</issue><spage>e0256220</spage><epage>e0256220</epage><pages>e0256220-e0256220</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. Here, we establish a reliable 1F5 based-icELISA that constitutes a promising diagnostic tool for control strategies and the prevention of ZIKV propagation.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>34403457</pmid><doi>10.1371/journal.pone.0256220</doi><tpages>e0256220</tpages><orcidid>https://orcid.org/0000-0002-8307-2100</orcidid><oa>free_for_read</oa></addata></record> |
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recordid | cdi_plos_journals_2562226311 |
source | MEDLINE; DOAJ Directory of Open Access Journals; Public Library of Science (PLoS) Journals Open Access; EZB-FREE-00999 freely available EZB journals; PubMed Central; Free Full-Text Journals in Chemistry |
subjects | Analysis Animals Antibodies, Monoclonal - biosynthesis Antibodies, Monoclonal - chemistry Antibodies, Monoclonal - isolation & purification Antibodies, Viral - biosynthesis Antibodies, Viral - chemistry Antibodies, Viral - isolation & purification Antigens, Viral - administration & dosage Antigens, Viral - genetics Antigens, Viral - immunology Binding, Competitive Biology and Life Sciences Chromatography Cloning, Molecular Congenital defects Cross-reaction Dengue fever Early Diagnosis Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - methods Enzyme-Linked Immunosorbent Assay - standards Enzymes Epitopes Escherichia coli - genetics Escherichia coli - metabolism Flaviviridae Gene Expression Global health Guillain-Barre syndrome Health risks HEK293 Cells Humans Hybridomas - chemistry Hybridomas - immunology Immunization Infections Male Markers Medicine and Health Sciences Mice Mice, Inbred BALB C Monoclonal antibodies NS1 protein Protein Multimerization Proteins Public health Recombinant Proteins - administration & dosage Recombinant Proteins - genetics Recombinant Proteins - immunology Reproducibility of Results Research and Analysis Methods Sensitivity and Specificity Vaccines Vector-borne diseases Viral Nonstructural Proteins - administration & dosage Viral Nonstructural Proteins - genetics Viral Nonstructural Proteins - immunology Viruses West Nile virus Zika virus Zika Virus - genetics Zika Virus - immunology Zika Virus Infection - diagnosis Zika Virus Infection - immunology Zika Virus Infection - virology |
title | Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection |
url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-19T11%3A08%3A30IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Development%20of%20a%20novel%20NS1%20competitive%20enzyme-linked%20immunosorbent%20assay%20for%20the%20early%20detection%20of%20Zika%20virus%20infection&rft.jtitle=PloS%20one&rft.au=Rold%C3%A1n,%20Julieta%20S&rft.date=2021-08-17&rft.volume=16&rft.issue=8&rft.spage=e0256220&rft.epage=e0256220&rft.pages=e0256220-e0256220&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0256220&rft_dat=%3Cgale_plos_%3EA672365757%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2562226311&rft_id=info:pmid/34403457&rft_galeid=A672365757&rft_doaj_id=oai_doaj_org_article_02b80f0c52a34fa79609d4a99d1db175&rfr_iscdi=true |