{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T15:09:48Z","timestamp":1780499388338,"version":"3.54.1"},"reference-count":56,"publisher":"American Society of Tropical Medicine and Hygiene","issue":"6","license":[{"start":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T00:00:00Z","timestamp":1780444800000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,6,3]]},"abstract":"<jats:title>ABSTRACT.<\/jats:title>\n                  <jats:p>\n                    Programs designed to eliminate\n                    <jats:italic>Onchocerca volvulus<\/jats:italic>\n                    transmission rely on measurement of Ov16-specific antibodies in children &lt;10 years old and molecular detection of parasites in blackfly vectors. Antibody testing can be conducted using rapid diagnostic tests or an ELISA, depending on location of testing and sample throughput needs. In previous efforts to establish anti-Ov16 testing in endemic country laboratories using existing tests, we encountered difficulties with assay reproducibility, reagent availability and shipping, and user friendliness. To address these challenges, we developed and validated an ELISA using Ov16 recombinant antigen expressed in a mammalian protein expression system that can be completed within 2 hours and does not require refrigeration during shipment. The assay was developed and validated using two distinct sets of dried blood spot samples that included defined positive, negative, and samples from infections with potential cross-reactivity. During development, the ELISA had 92.98% (95% CI = 88.28\u201397.68%) sensitivity and a specificity of 98.50% (95% CI = 93.80\u2013100%). The cutoff point established during development was then applied to the validation set of specimens, which demonstrated a sensitivity of 89.47% (95% CI = 84.77\u201394.17%) and 98.32% (95% CI = 93.62\u2013100%) specificity. When repeated over 4 days by 2 different operators, all replicates yielded reproducible results within a coefficient of variance of 20%. In addition, we performed stability testing on assay reagents to evaluate their suitability for shipping at ambient temperatures. The resulting assay will be useful for monitoring anti-Ov16 prevalence in endemic country laboratories.\n                  <\/jats:p>","DOI":"10.4269\/ajtmh.25-0725","type":"journal-article","created":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T17:11:15Z","timestamp":1775581875000},"page":"1172-1179","source":"Crossref","is-referenced-by-count":0,"title":["Development and Validation of an ELISA to Detect Antibody to Onchocerca volvulus Infection Using Mammalian Expressed Recombinant Ov16"],"prefix":"10.4269","volume":"114","author":[{"given":"Sylvia","family":"Ossai","sequence":"first","affiliation":[{"name":"CDC Foundation, Atlanta, Georgia;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong","family":"Wang","sequence":"additional","affiliation":[{"name":"Synergy America, Inc., Duluth, Georgia, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eric S.","family":"Elder","sequence":"additional","affiliation":[{"name":"Laboratory Science and Diagnostics Branch, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, Georgia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karana","family":"Wickens","sequence":"additional","affiliation":[{"name":"Laboratory Science and Diagnostics Branch, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, Georgia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kimberly Y.","family":"Won","sequence":"additional","affiliation":[{"name":"Laboratory Science and Diagnostics Branch, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, Georgia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. Evan","family":"Secor","sequence":"additional","affiliation":[{"name":"Laboratory Science and Diagnostics Branch, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, Georgia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sukwan","family":"Handali","sequence":"additional","affiliation":[{"name":"Laboratory Science and Diagnostics Branch, Division of Parasitic Diseases and Malaria, Centers for Disease Control and Prevention, Atlanta, Georgia, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"2795","reference":[{"key":"b1","series-title":"Gates Open Res","first-page":"1545","article-title":"The World Health Organization 2030 goals for onchocerciasis: Insights and perspectives from mathematical modelling: NTD Modelling Consortium Onchocerciasis Group","volume":"3","year":"2019"},{"key":"b2","series-title":"Clin Infect Dis","first-page":"S165","article-title":"Scaling-down mass ivermectin treatment for onchocerciasis elimination: Modeling the impact of the geographical unit for decision making","volume":"72","author":"Stolk","year":"2021"},{"key":"b3","series-title":"Front Trop Dis","first-page":"986884","article-title":"Onchocerciasis: Current knowledge and future goals","volume":"3","author":"Frallonardo","year":"2022"},{"key":"b4","series-title":"Open Forum Infect Dis","first-page":"ofaa206","article-title":"A second population-based cohort study in Cameroon confirms the temporal relationship between onchocerciasis and epilepsy","volume":"7","author":"Chesnais","year":"2020"},{"key":"b5","series-title":"Glob Ment Health (Camb)","first-page":"e26","article-title":"Nodding syndrome research, lessons learned from the NSETHIO project","volume":"6","author":"Geelhand de Merxem","year":"2019"},{"key":"b6","article-title":"Guidelines for Stopping Mass Drug Administration and Verifying Elimination of Human Onchocerciasis: Criteria and Procedures.","year":"2016"},{"key":"b7","series-title":"J Infect Dis","first-page":"282","article-title":"Detection of Onchocerca volvulus infection by O-150 polymerase chain reaction analysis of skin scratches","volume":"178","author":"To\u00e9","year":"1998"},{"key":"b8","series-title":"Mol Biochem Parasitol","first-page":"135","article-title":"Identification of an Onchocerca volvulus cDNA encoding a low-molecular-weight antigen uniquely recognized by onchocerciasis patient sera","volume":"39","author":"Lobos","year":"1990"},{"key":"b9","series-title":"Science","first-page":"1603","article-title":"An immunogenic Onchocerca volvulus antigen: A specific and early marker of infection","volume":"251","author":"Lobos","year":"1991"},{"key":"b10","series-title":"Am J Trop Med Hyg","first-page":"1041","article-title":"Evaluation of an OV-16 IgG4 enzyme-linked immunosorbent assay in humans and its application to determine the dynamics of antibody responses in a non-human primate model of Onchocerca volvulus infection","volume":"99","author":"Cama","year":"2018"},{"key":"b11","series-title":"PLoS Negl Trop Dis","first-page":"e0004292","article-title":"A recombinant positive control for serology diagnostic tests supporting elimination of Onchocerca volvulus.","volume":"10","author":"Golden","year":"2016"},{"key":"b12","series-title":"Parasite Epidemiol Control","first-page":"e00352","article-title":"Field sensitivity and specificity of the SD BIOLINE onchocerciasis IgG4 Rapid Diagnostic Test in children <10 years old from endemic areas in Burkina Faso","volume":"25","author":"Niki\u00e8ma","year":"2024"},{"key":"b13","series-title":"Am J Trop Med Hyg","first-page":"749","article-title":"Operational performance of the Onchocerca volvulus \u201cOEPA\u201d Ov16 ELISA serological assay in mapping, guiding decisions to stop mass drug administration, and posttreatment surveillance surveys","volume":"99","author":"Richards","year":"2018"},{"key":"b14","series-title":"Am J Trop Med Hyg","first-page":"779","article-title":"Evaluation of the diagnostic performance of Onchocerca volvulus linear epitopes in a peptide enzyme-linked immunosorbent assay","volume":"98","author":"Lagatie","year":"2018"},{"key":"b15","series-title":"Am J Trop Med Hyg","first-page":"339","article-title":"Serosurveillance to monitor onchocerciasis elimination: The Ugandan experience","volume":"90","author":"Oguttu","year":"2014"},{"key":"b16","series-title":"PLoS Negl Trop Dis","first-page":"e0010682","article-title":"Onchocerciasis: Target product profiles of in\u00a0vitro diagnostics to support onchocerciasis elimination mapping and mass drug administration stopping decisions","volume":"16","author":"Biamonte","year":"2022"},{"key":"b17","series-title":"PLoS Negl Trop Dis","first-page":"e0011505","article-title":"Correction: Onchocerciasis: Target product profiles of in\u00a0vitro diagnostics to support onchocerciasis elimination mapping and mass drug administration stopping decisions","volume":"17","author":"Biamonte","year":"2023"},{"key":"b18","series-title":"PLoS Negl Trop Dis","first-page":"e1941","article-title":"Longitudinal monitoring of the development of antifilarial antibodies and acquisition of Wuchereria bancrofti in a highly endemic area of Haiti","volume":"6","author":"Hamlin","year":"2012"},{"key":"b19","series-title":"J Immunol","first-page":"4001","article-title":"Guatemalan human onchocerciasis. II. Evidence for IgG3 involvement in acquired immunity to Onchocerca volvulus and identification of possible immune-associated antigens","volume":"146","author":"Boyer","year":"1991"},{"key":"b20","series-title":"J Immunol","first-page":"3993","article-title":"Guatemalan human onchocerciasis. I. Systematic analysis of patient populations, nodular antigens, and specific isotypic reactions","volume":"146","author":"Tsang","year":"1991"},{"key":"b21","series-title":"Am J Trop Med Hyg","first-page":"666","article-title":"Characterizing reactivity to Onchocerca volvulus antigens in multiplex bead assays","volume":"97","author":"Feeser","year":"2017"},{"key":"b22","series-title":"PLoS Negl Trop Dis","first-page":"e0008008","article-title":"Post-elimination surveillance in formerly onchocerciasis endemic focus in Southern Mexico","volume":"14","author":"Fern\u00e1ndez-Santos","year":"2020"},{"key":"b23","series-title":"BMC Bioinformatics","first-page":"77","article-title":"pROC: An open-source package for R and S+ to analyze and compare ROC curves","volume":"12","author":"Robin","year":"2011"},{"key":"b24","article-title":"Onchocerciasis: Diagnostic Target Product Profile to Support Preventive Chemotherapy.","year":"2021"},{"key":"b25","series-title":"PLoS Negl Trop Dis","first-page":"e0008933","article-title":"Time for a diagnostic sea-change: Rethinking neglected tropical disease diagnostics to achieve elimination","volume":"14","author":"Gass","year":"2020"},{"key":"b26","series-title":"Parasit Vectors","first-page":"176","article-title":"Expression of Ascaris lumbricoides putative virulence-associated genes when infecting a human host","volume":"14","author":"Mohd-Shaharuddin","year":"2021"},{"key":"b27","series-title":"Int Health","first-page":"i20","article-title":"Diagnostics for onchocerciasis in the era of elimination","volume":"10","author":"Unnasch","year":"2018"},{"key":"b28","series-title":"Parasite Immunol","first-page":"89","article-title":"Immuno-epidemiology of Ascaris lumbricoides infection in a high transmission community: Antibody responses and their impact on current and future infection intensity","volume":"27","author":"King","year":"2005"},{"key":"b1","series-title":"Gates Open Res","first-page":"1545","article-title":"The World Health Organization 2030 goals for onchocerciasis: Insights and perspectives from mathematical modelling: NTD Modelling Consortium Onchocerciasis Group","volume":"3","year":"2019"},{"key":"b2","series-title":"Clin Infect Dis","first-page":"S165","article-title":"Scaling-down mass ivermectin treatment for onchocerciasis elimination: Modeling the impact of the geographical unit for decision making","volume":"72","author":"Stolk","year":"2021"},{"key":"b3","series-title":"Front Trop Dis","first-page":"986884","article-title":"Onchocerciasis: Current knowledge and future goals","volume":"3","author":"Frallonardo","year":"2022"},{"key":"b4","series-title":"Open Forum Infect Dis","first-page":"ofaa206","article-title":"A second population-based cohort study in Cameroon confirms the temporal relationship between onchocerciasis and epilepsy","volume":"7","author":"Chesnais","year":"2020"},{"key":"b5","series-title":"Glob Ment Health (Camb)","first-page":"e26","article-title":"Nodding syndrome research, lessons learned from the NSETHIO project","volume":"6","author":"Geelhand de Merxem","year":"2019"},{"key":"b6","article-title":"Guidelines for Stopping Mass Drug Administration and Verifying Elimination of Human Onchocerciasis: Criteria and Procedures.","year":"2016"},{"key":"b7","series-title":"J Infect Dis","first-page":"282","article-title":"Detection of Onchocerca volvulus infection by O-150 polymerase chain reaction analysis of skin scratches","volume":"178","author":"To\u00e9","year":"1998"},{"key":"b8","series-title":"Mol Biochem Parasitol","first-page":"135","article-title":"Identification of an Onchocerca volvulus cDNA encoding a low-molecular-weight antigen uniquely recognized by onchocerciasis patient sera","volume":"39","author":"Lobos","year":"1990"},{"key":"b9","series-title":"Science","first-page":"1603","article-title":"An immunogenic Onchocerca volvulus antigen: A specific and early marker of infection","volume":"251","author":"Lobos","year":"1991"},{"key":"b10","series-title":"Am J Trop Med Hyg","first-page":"1041","article-title":"Evaluation of an OV-16 IgG4 enzyme-linked immunosorbent assay in humans and its application to determine the dynamics of antibody responses in a non-human primate model of Onchocerca volvulus infection","volume":"99","author":"Cama","year":"2018"},{"key":"b11","series-title":"PLoS Negl Trop Dis","first-page":"e0004292","article-title":"A recombinant positive control for serology diagnostic tests supporting elimination of Onchocerca volvulus.","volume":"10","author":"Golden","year":"2016"},{"key":"b12","series-title":"Parasite Epidemiol Control","first-page":"e00352","article-title":"Field sensitivity and specificity of the SD BIOLINE onchocerciasis IgG4 Rapid Diagnostic Test in children <10 years old from endemic areas in Burkina Faso","volume":"25","author":"Niki\u00e8ma","year":"2024"},{"key":"b13","series-title":"Am J Trop Med Hyg","first-page":"749","article-title":"Operational performance of the Onchocerca volvulus \u201cOEPA\u201d Ov16 ELISA serological assay in mapping, guiding decisions to stop mass drug administration, and posttreatment surveillance surveys","volume":"99","author":"Richards","year":"2018"},{"key":"b14","series-title":"Am J Trop Med Hyg","first-page":"779","article-title":"Evaluation of the diagnostic performance of Onchocerca volvulus linear epitopes in a peptide enzyme-linked immunosorbent assay","volume":"98","author":"Lagatie","year":"2018"},{"key":"b15","series-title":"Am J Trop Med Hyg","first-page":"339","article-title":"Serosurveillance to monitor onchocerciasis elimination: The Ugandan experience","volume":"90","author":"Oguttu","year":"2014"},{"key":"b16","series-title":"PLoS Negl Trop Dis","first-page":"e0010682","article-title":"Onchocerciasis: Target product profiles of in\u00a0vitro diagnostics to support onchocerciasis elimination mapping and mass drug administration stopping decisions","volume":"16","author":"Biamonte","year":"2022"},{"key":"b17","series-title":"PLoS Negl Trop Dis","first-page":"e0011505","article-title":"Correction: Onchocerciasis: Target product profiles of in\u00a0vitro diagnostics to support onchocerciasis elimination mapping and mass drug administration stopping decisions","volume":"17","author":"Biamonte","year":"2023"},{"key":"b18","series-title":"PLoS Negl Trop Dis","first-page":"e1941","article-title":"Longitudinal monitoring of the development of antifilarial antibodies and acquisition of Wuchereria bancrofti in a highly endemic area of Haiti","volume":"6","author":"Hamlin","year":"2012"},{"key":"b19","series-title":"J Immunol","first-page":"4001","article-title":"Guatemalan human onchocerciasis. II. Evidence for IgG3 involvement in acquired immunity to Onchocerca volvulus and identification of possible immune-associated antigens","volume":"146","author":"Boyer","year":"1991"},{"key":"b20","series-title":"J Immunol","first-page":"3993","article-title":"Guatemalan human onchocerciasis. I. Systematic analysis of patient populations, nodular antigens, and specific isotypic reactions","volume":"146","author":"Tsang","year":"1991"},{"key":"b21","series-title":"Am J Trop Med Hyg","first-page":"666","article-title":"Characterizing reactivity to Onchocerca volvulus antigens in multiplex bead assays","volume":"97","author":"Feeser","year":"2017"},{"key":"b22","series-title":"PLoS Negl Trop Dis","first-page":"e0008008","article-title":"Post-elimination surveillance in formerly onchocerciasis endemic focus in Southern Mexico","volume":"14","author":"Fern\u00e1ndez-Santos","year":"2020"},{"key":"b23","series-title":"BMC Bioinformatics","first-page":"77","article-title":"pROC: An open-source package for R and S+ to analyze and compare ROC curves","volume":"12","author":"Robin","year":"2011"},{"key":"b24","article-title":"Onchocerciasis: Diagnostic Target Product Profile to Support Preventive Chemotherapy.","year":"2021"},{"key":"b25","series-title":"PLoS Negl Trop Dis","first-page":"e0008933","article-title":"Time for a diagnostic sea-change: Rethinking neglected tropical disease diagnostics to achieve elimination","volume":"14","author":"Gass","year":"2020"},{"key":"b26","series-title":"Parasit Vectors","first-page":"176","article-title":"Expression of Ascaris lumbricoides putative virulence-associated genes when infecting a human host","volume":"14","author":"Mohd-Shaharuddin","year":"2021"},{"key":"b27","series-title":"Int Health","first-page":"i20","article-title":"Diagnostics for onchocerciasis in the era of elimination","volume":"10","author":"Unnasch","year":"2018"},{"key":"b28","series-title":"Parasite Immunol","first-page":"89","article-title":"Immuno-epidemiology of Ascaris lumbricoides infection in a high transmission community: Antibody responses and their impact on current and future infection intensity","volume":"27","author":"King","year":"2005"}],"container-title":["The American Journal of Tropical Medicine and Hygiene"],"original-title":[],"link":[{"URL":"https:\/\/www.ajtmh.org\/view\/journals\/tpmd\/114\/6\/article-p1172.xml","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.ajtmh.org\/downloadpdf\/journals\/tpmd\/114\/6\/article-p1172.xml","content-type":"text\/html","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/www.ajtmh.org\/downloadpdf\/journals\/tpmd\/114\/6\/article-p1172.xml","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T14:16:10Z","timestamp":1780496170000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ajtmh.org\/view\/journals\/tpmd\/114\/6\/article-p1172.xml"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,3]]},"references-count":56,"journal-issue":{"issue":"6"},"URL":"https:\/\/doi.org\/10.4269\/ajtmh.25-0725","relation":{},"ISSN":["0002-9637","1476-1645"],"issn-type":[{"value":"0002-9637","type":"print"},{"value":"1476-1645","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,3]]}}}