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Although symptoms are typically mild and unspecific, Zika\u2019s major impact occurs during pregnancy, generating a congenital syndrome. Serology plays a key role in its diagnosis. However, its use is limited due to the uncertainty caused by the cross-reaction of antibodies elicited in response to other flavivirus infections when tested in direct immunoassays. Using a panel of previously generated anti-Zika non-structural protein 1 (NS1) nanobodies, a set was selected that only recognizes epitopes present in Zika and is immunogenic to humans. A proper arrangement of these nanobodies was made and conditions were optimized in order to develop a novel serology assay. This new ELISA relies on the inhibition of the binding of a set of selected nanobodies to Zika-immobilized NS1 when previously incubated with Zika convalescent sera. Using the developed blocking of binding assay, it was possible to discriminate between Zika-specific and cross-reactive antibodies in serum samples from infections with Zika and other flaviviruses.<\/jats:p>","DOI":"10.3390\/tropicalmed8010055","type":"journal-article","created":{"date-parts":[[2023,1,11]],"date-time":"2023-01-11T03:40:35Z","timestamp":1673408435000},"page":"55","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Nanobody-Based Blocking of Binding ELISA for the Detection of Anti-NS1 Zika-Virus-Specific Antibodies in Convalescent Patients"],"prefix":"10.3390","volume":"8","author":[{"given":"Triana","family":"Delfin-Riela","sequence":"first","affiliation":[{"name":"C\u00e1tedra de Inmunolog\u00eda, DEPBIO, Facultad de Qu\u00edmica, Instituto de Higiene, UDELAR, Montevideo 11600, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8902-5532","authenticated-orcid":false,"given":"Mart\u00edn A.","family":"Rossotti","sequence":"additional","affiliation":[{"name":"C\u00e1tedra de Inmunolog\u00eda, DEPBIO, Facultad de Qu\u00edmica, Instituto de Higiene, UDELAR, Montevideo 11600, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9475-1590","authenticated-orcid":false,"given":"Giada","family":"Mattiuzzo","sequence":"additional","affiliation":[{"name":"Division of Virology, National Institute for Biological Standards and Control (NIBSC)-MHRA, Hertfordshire EN6 3QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C\u00e9sar","family":"Echaides","sequence":"additional","affiliation":[{"name":"Parque Lecoq, IMM, Montevideo 11300, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1824-0611","authenticated-orcid":false,"given":"Gualberto","family":"Gonz\u00e1lez-Sapienza","sequence":"additional","affiliation":[{"name":"C\u00e1tedra de Inmunolog\u00eda, DEPBIO, Facultad de Qu\u00edmica, Instituto de Higiene, UDELAR, Montevideo 11600, Uruguay"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/0035-9203(52)90042-4","article-title":"Zika virus (I). Isolations and serological specificity","volume":"46","author":"Dick","year":"1952","journal-title":"Trans. R. Soc. Trop. Med. Hyg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"116","DOI":"10.3855\/jidc.8217","article-title":"Zika virus outbreak in Brazil","volume":"10","author":"Heukelbach","year":"2016","journal-title":"J. Infect. Dev. Ctries."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1126\/science.aaf5036","article-title":"Zika virus in the Americas: Early epidemiological and genetic findings","volume":"352","author":"Faria","year":"2016","journal-title":"Science"},{"key":"ref_4","first-page":"1085","article-title":"Zika virus, French polynesia, South pacific, 2013","volume":"20","author":"Roche","year":"2014","journal-title":"Emerg. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"359","DOI":"10.3201\/eid2102.141363","article-title":"Potential sexual transmission of Zika virus","volume":"21","author":"Musso","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"951","DOI":"10.1056\/NEJMoa1600651","article-title":"Zika Virus Associated with Microcephaly","volume":"374","author":"Mlakar","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1001\/jamapediatrics.2016.3982","article-title":"Characterizing the Pattern of Anomalies in Congenital Zika Syndrome for Pediatric Clinicians","volume":"171","author":"Moore","year":"2017","journal-title":"JAMA Pediatr."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"475","DOI":"10.15585\/mmwr.mm6518e2","article-title":"Comparison of Test Results for Zika Virus RNA in Urine, Serum, and Saliva Specimens from Persons with Travel-Associated Zika Virus Disease\u2014Florida, 2016","volume":"65","author":"Bingham","year":"2016","journal-title":"MMWR Morb. Mortal. Wkly. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1038\/nsmb.3213","article-title":"Zika virus NS1 structure reveals diversity of electrostatic surfaces among flaviviruses","volume":"23","author":"Song","year":"2016","journal-title":"Nat. Struct. Mol. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.1038\/ni.3515","article-title":"Dengue virus sero-cross-reactivity drives antibody-dependent enhancement of infection with zika virus","volume":"17","author":"Dejnirattisai","year":"2016","journal-title":"Nat. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7852","DOI":"10.1073\/pnas.1607931113","article-title":"Human antibody responses after dengue virus infection are highly cross-reactive to Zika virus","volume":"113","author":"Priyamvada","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.3201\/eid2501.180605","article-title":"Dengue Virus IgM Serotyping by ELISA with Recombinant Mutant Envelope Proteins","volume":"25","author":"Rockstroh","year":"2019","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1080\/21505594.2019.1656503","article-title":"Host and viral mechanisms of congenital Zika syndrome","volume":"10","author":"Liang","year":"2019","journal-title":"Virulence"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/(SICI)1096-9071(199901)57:1<1::AID-JMV1>3.0.CO;2-T","article-title":"Antibody responses to an immunodominant nonstructural 1 synthetic peptide in patients with dengue fever and dengue hemorrhagic fever","volume":"57","author":"Huang","year":"1999","journal-title":"J. Med. Virol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2489","DOI":"10.1128\/JCM.42.6.2489-2494.2004","article-title":"Dengue virus serotyping based on envelope and membrane and nonstructural protein NS1 serotype-specific capture immunoglobulin M enzyme-linked immunosorbent assays","volume":"42","author":"Shu","year":"2004","journal-title":"J. Clin. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1477","DOI":"10.1002\/jmv.24789","article-title":"Cross reactivity of commercial anti-dengue immunoassays in patients with acute Zika virus infection","volume":"89","author":"Felix","year":"2017","journal-title":"J. Med. Virol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"105201","DOI":"10.1016\/j.actatropica.2019.105201","article-title":"Serological tests reveal significant cross-reactive human antibody responses to Zika and Dengue viruses in the Mexican population","volume":"201","author":"Zaidi","year":"2020","journal-title":"Acta Trop."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"8384","DOI":"10.1073\/pnas.1704984114","article-title":"Antibody-based assay discriminates Zika virus infection from other flaviviruses","volume":"114","author":"Balmaseda","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"446","DOI":"10.1038\/363446a0","article-title":"Naturally occurring antibodies devoid of light chains","volume":"363","author":"Atarhouch","year":"1993","journal-title":"Nature"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"775","DOI":"10.1146\/annurev-biochem-063011-092449","article-title":"Nanobodies: Natural single-domain antibodies","volume":"82","author":"Muyldermans","year":"2013","journal-title":"Annu. Rev. Biochem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"977","DOI":"10.3389\/fimmu.2017.00977","article-title":"Single-Domain Antibodies As Versatile Affinity Reagents for Analytical and Diagnostic Applications","volume":"8","author":"Rossotti","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1016\/S0167-4838(99)00030-8","article-title":"Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies","volume":"1431","author":"Frenken","year":"1999","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1007\/s00216-019-02246-7","article-title":"A nanobody-based test for highly sensitive detection of hemoglobin in fecal samples","volume":"412","author":"Rossotti","year":"2020","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"11907","DOI":"10.1021\/acs.analchem.5b03561","article-title":"Method for Sorting and Pairwise Selection of Nanobodies for the Development of Highly Sensitive Sandwich Immunoassays","volume":"87","author":"Rossotti","year":"2015","journal-title":"Anal. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Delfin-Riela, T., Rossotti, M., Alvez-Rosado, R., Leizagoyen, C., and Gonz\u00e1lez-Sapienza, G. (2020). Highly Sensitive Detection of Zika Virus Nonstructural Protein 1 in Serum Samples by a Two-Site Nanobody ELISA. Biomolecules, 10.","DOI":"10.3390\/biom10121652"},{"key":"ref_26","unstructured":"Morillas, P.P., Terra, A.I., Uribe, C., Mastromonaco, G., Prieto, J.L., and Torres, M. (2016). Gu\u00eda Eurachem: La Adecuaci\u00f3n al uso de los M\u00e9todos Anal\u00edticos\u2014Una Gu\u00eda de Laboratorio Para la Validaci\u00f3n de M\u00e9todos y Temas Relacionados, EURACHEM, Eurolab Espa\u00f1aT."}],"container-title":["Tropical Medicine and Infectious Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2414-6366\/8\/1\/55\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:06:02Z","timestamp":1760119562000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2414-6366\/8\/1\/55"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,10]]},"references-count":26,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["tropicalmed8010055"],"URL":"https:\/\/doi.org\/10.3390\/tropicalmed8010055","relation":{},"ISSN":["2414-6366"],"issn-type":[{"value":"2414-6366","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,10]]}}}