{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T18:31:07Z","timestamp":1783017067271,"version":"3.54.6"},"reference-count":23,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T00:00:00Z","timestamp":1782950400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100016302","name":"Coalition for Epidemic Preparedness Innovations","doi-asserted-by":"publisher","award":["276869"],"award-info":[{"award-number":["276869"]}],"id":[{"id":"10.13039\/100016302","id-type":"DOI","asserted-by":"publisher"}]},{"name":"EDCTP2","award":["RIA2019LV-3053"],"award-info":[{"award-number":["RIA2019LV-3053"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Lassa fever is a severe, often fatal febrile illness endemic to West Africa caused by Lassa virus (LASV), with different virus lineages predominating across West African countries. The viral nucleoprotein (NP) is a target antigen for serological assays to identify previous exposure to LASV. To our knowledge, there is no commercially available assay that reliably quantifies anti-LASV-NP IgG antibodies in human serum. We report the development and qualification of an ELISA designed to detect and quantify anti-LASV-NP IgG in human serum samples. Following assay optimization, performance was assessed through assay qualification at clinical trial laboratories within Ghana. Assay positivity criteria, lower limit of detection, upper and lower limits of quantification, inter-assay precision, selectivity and dilutional linearity were determined. A new reference standard prepared from pooled sera from donors in endemic Lassa fever regions was established and calibrated to the first WHO international standard for LASV antibodies. One ELISA assay utilizing lineage IV LASV-NP was applicable for detection of anti-LASV-NP IgG antibodies in serum samples from different West African countries where either LASV lineages I, II, III and IV predominate. The ELISA remained selective in hemolysed serum samples with minimal loss of signal across repeated sample freeze-thaw cycles. Crucially, the developed ELISA was fully concordant with a now discontinued commercially available ELISA kit for quantification of anti-LASV-NP antibodies. Our anti-LASV-NP IgG ELISA was shown to reliably measure anti-LASV-NP IgG levels in human serum. Establishing and conducting this assay within West Africa represents an essential step towards strengthening LASV epidemiology research and supporting urgently needed development of a vaccine to prevent Lassa Fever.<\/jats:p>","DOI":"10.1371\/journal.pone.0340568","type":"journal-article","created":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T17:32:16Z","timestamp":1783013536000},"page":"e0340568","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and qualification of an enzyme-linked immunosorbent assay to detect human serum immunoglobulin G reactive to multiple lineages of Lassa virus nucleoprotein"],"prefix":"10.1371","volume":"21","author":[{"given":"Heejin","family":"Yun","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Faith","family":"Sigei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana Yaa Asiedua","family":"Appiah","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte Naa Odey","family":"Quaye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cecilia Adwoa Biaa","family":"Yankey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6423-1885","authenticated-orcid":true,"given":"Eric","family":"Kyei-Baafour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1251-934X","authenticated-orcid":true,"given":"Peter","family":"Hayes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3416-7362","authenticated-orcid":true,"given":"Arianna","family":"Marini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert T.","family":"Bailer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0569-6049","authenticated-orcid":true,"given":"Marija","family":"Zaric","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5483-9985","authenticated-orcid":true,"given":"Kwadwo Asamoah","family":"Kusi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,7,2]]},"reference":[{"issue":"2","key":"pone.0340568.ref001","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1038\/s41579-022-00789-8","article-title":"Lassa fever - the road ahead","volume":"21","author":"R Garry","year":"2023","journal-title":"Nat Rev Microbiol"},{"issue":"3","key":"pone.0340568.ref002","doi-asserted-by":"crossref","first-page":"599","DOI":"10.3201\/eid2403.171905","article-title":"New lineage of Lassa virus, Togo, 2016","volume":"24","author":"S Whitmer","year":"2018","journal-title":"Emerg Infect Dis"},{"issue":"1","key":"pone.0340568.ref003","doi-asserted-by":"crossref","first-page":"1761","DOI":"10.1080\/22221751.2020.1796528","article-title":"Lassa fever in Benin: description of the 2014 and 2016 epidemics and genetic characterization of a new Lassa virus","volume":"9","author":"A Yadouleton","year":"2020","journal-title":"Emerg Microbes Infect"},{"issue":"1","key":"pone.0340568.ref004","doi-asserted-by":"crossref","first-page":"146","DOI":"10.3390\/v15010146","article-title":"Combating Lassa Fever in West African Sub-Region: Progress, Challenges, and Future Perspectives","volume":"15","author":"C Aloke","year":"2023","journal-title":"Viruses"},{"key":"pone.0340568.ref005","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41541-018-0049-5","article-title":"Baseline mapping of Lassa fever virology, epidemiology and vaccine research and development","volume":"3","author":"HJ Hallam","year":"2018","journal-title":"NPJ Vaccines"},{"key":"pone.0340568.ref006","unstructured":"WHO. Pathogens prioritization: a scientific framework for epidemic and pandemic research preparedness. 2024. https:\/\/www.who.int\/publications\/m\/item\/pathogens-prioritization-a-scientific-framework-for-epidemic-and-pandemic-research-preparedness"},{"issue":"10","key":"pone.0340568.ref007","doi-asserted-by":"crossref","first-page":"5458","DOI":"10.1128\/JVI.78.10.5458-5465.2004","article-title":"Properties of replication-competent vesicular stomatitis virus vectors expressing glycoproteins of filoviruses and arenaviruses","volume":"78","author":"M Garbutt","year":"2004","journal-title":"J Virol"},{"issue":"3","key":"pone.0340568.ref008","doi-asserted-by":"crossref","DOI":"10.1128\/jvi.01627-23","article-title":"Translational success of fundamental virology: a VSV-vectored Ebola vaccine","volume":"98","author":"EM Anderson","year":"2024","journal-title":"J Virol"},{"key":"pone.0340568.ref009","volume-title":"Ebola Vaccine Product Information","author":"CDC","year":"2024"},{"issue":"16","key":"pone.0340568.ref010","doi-asserted-by":"crossref","first-page":"360","DOI":"10.15585\/mmwr.mm7316a1","article-title":"Use of Ebola Vaccines - Worldwide, 2021-2023","volume":"73","author":"R Kallay","year":"2024","journal-title":"MMWR Morb Mortal Wkly Rep"},{"issue":"3","key":"pone.0340568.ref011","doi-asserted-by":"crossref","DOI":"10.3390\/vaccines9030190","article-title":"Development of pandemic vaccines: ERVEBO case study","volume":"9","author":"J Wolf","year":"2021","journal-title":"Vaccines"},{"issue":"12","key":"pone.0340568.ref012","doi-asserted-by":"crossref","DOI":"10.1371\/journal.ppat.1010078","article-title":"Current state of Ebola virus vaccines: A snapshot","volume":"17","author":"C Woolsey","year":"2021","journal-title":"PLoS Pathog"},{"issue":"18","key":"pone.0340568.ref013","doi-asserted-by":"crossref","first-page":"1807","DOI":"10.1056\/NEJMoa2501073","article-title":"Safety and Immunogenicity of an rVSV Lassa Fever Vaccine Candidate","volume":"393","author":"E Malkin","year":"2025","journal-title":"N Engl J Med"},{"issue":"10","key":"pone.0340568.ref014","doi-asserted-by":"crossref","first-page":"2031","DOI":"10.3390\/v4102031","article-title":"Pathogenesis of Lassa fever","volume":"4","author":"NE Yun","year":"2012","journal-title":"Viruses"},{"issue":"4","key":"pone.0340568.ref015","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1016\/j.cell.2015.07.020","article-title":"Clinical Sequencing Uncovers Origins and Evolution of Lassa Virus","volume":"162","author":"KG Andersen","year":"2015","journal-title":"Cell"},{"issue":"15","key":"pone.0340568.ref016","doi-asserted-by":"crossref","first-page":"6992","DOI":"10.1128\/JVI.74.15.6992-7004.2000","article-title":"Genetic diversity among Lassa virus strains","volume":"74","author":"MD Bowen","year":"2000","journal-title":"J Virol"},{"issue":"9","key":"pone.0340568.ref017","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1128\/CVI.00101-07","article-title":"Development of Recombinant Nucleoprotein-Based Diagnostic Systems for Lassa Fever","volume":"14","author":"M Saijo","year":"2007","journal-title":"Clin Vaccine Immunol"},{"key":"pone.0340568.ref018","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1186\/1743-422X-7-279","article-title":"Lassa virus-like particles displaying all major immunological determinants as a vaccine candidate for Lassa hemorrhagic fever","volume":"7","author":"LM Branco","year":"2010","journal-title":"Virol J"},{"issue":"2","key":"pone.0340568.ref019","doi-asserted-by":"crossref","first-page":"130","DOI":"10.4314\/ajcem.v24i2.3","article-title":"A review of the recent advances on Lassa fever with special reference to molecular epidemiology and progress in vaccine development","volume":"24","author":"N Medugu","year":"2023","journal-title":"Afr J Clin Exp Micro"},{"issue":"3","key":"pone.0340568.ref020","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pntd.0009255","article-title":"Boosting understanding of Lassa Fever virus epidemiology: Field testing a novel assay to identify past Lassa Fever virus infection in blood and oral fluids of survivors and unexposed controls in Sierra Leone","volume":"15","author":"O Akpogheneta","year":"2021","journal-title":"PLoS Negl Trop Dis"},{"issue":"8423","key":"pone.0340568.ref021","doi-asserted-by":"crossref","first-page":"250","DOI":"10.1016\/S0140-6736(85)91029-3","article-title":"Early diagnosis of human Lassa fever by ELISA detection of antigen and antibody","volume":"1","author":"PB Jahrling","year":"1985","journal-title":"Lancet"},{"issue":"3","key":"pone.0340568.ref022","doi-asserted-by":"crossref","first-page":"210","DOI":"10.1002\/jmv.1890400308","article-title":"Lassa virus activity in Guinea: distribution of human antiviral antibody defined using enzyme-linked immunosorbent assay with recombinant antigen","volume":"40","author":"IS Lukashevich","year":"1993","journal-title":"J Med Virol"},{"issue":"6","key":"pone.0340568.ref023","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1111\/tmi.13876","article-title":"Lassa fever vaccine candidates: A scoping review of vaccine clinical trials","volume":"28","author":"G Sulis","year":"2023","journal-title":"Trop Med Int Health"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0340568","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,2]],"date-time":"2026-07-02T17:32:33Z","timestamp":1783013553000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0340568"}},"subtitle":[],"editor":[{"given":"Masahiro","family":"Kajihara","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,7,2]]},"references-count":23,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7,2]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0340568","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,2]]}}}