TYPE Original Research PUBLISHED 01 February 2023 DOI 10.3389/fvets.2023.1120367 OPEN ACCESS EDITED BY Fernando A. Osorio, University of Nebraska System, United States REVIEWED BY Korakrit Poonsuk, University of Nebraska-Lincoln, United States Jitendra Kumar Biswal, International Centre for Foot-and-Mouth Disease, India *CORRESPONDENCE Ming Yang ming.yang@inspection.gc.ca SPECIALTY SECTION This article was submitted to Veterinary Infectious Diseases, a section of the journal Frontiers in Veterinary Science RECEIVED 16 December 2022 ACCEPTED 13 January 2023 PUBLISHED 01 February 2023 CITATION Zhu W, Pickering B, Smith G, Pinette M, Truong T, Babiuk S, Kobasa D, Banadyga L and Yang M (2023) Development and laboratory evaluation of a competitive ELISA for serodiagnosis of Nipah and Hendra virus infection using recombinant Nipah glycoproteins and a monoclonal antibody. Front. Vet. Sci. 10:1120367. doi: 10.3389/fvets.2023.1120367 COPYRIGHT © 2023 Zhu, Pickering, Smith, Pinette, Truong, Babiuk, Kobasa, Banadyga and Yang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Development and laboratory evaluation of a competitive ELISA for serodiagnosis of Nipah and Hendra virus infection using recombinant Nipah glycoproteins and a monoclonal antibody Wenjun Zhu 1 , Bradley Pickering 1,2,3 , Greg Smith 1 , Mathieu Pinette 1 , Thang Truong 4 , Shawn Babiuk 1,5 , Darwyn Kobasa 2,4 , Logan Banadyga 2,4 and Ming Yang 1 * 1 National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB, Canada, 2 Department of Medical Microbiology and Infectious Diseases, University of Manitoba, Winnipeg, MB, Canada, 3 Department of Veterinary Microbiology and Preventative Medicine, College of Veterinary Medicine, Iowa State University, Ames, IA, United States, 4 National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, MB, Canada, 5 Department of Immunology, University of Manitoba, Winnipeg, MB, Canada Introduction: Nipah virus (NiV) and Hendra virus (HeV), of the genus Henipavirus, family Paramyxoviridae, are classified as Risk Group 4 (RG4) pathogens that cause respiratory disease in pigs and acute/febrile encephalitis in humans with high mortality. Methods: A competitive enzyme–linked immunosorbent assay (cELISA) using a monoclonal antibody (mAb) and recombinant NiV glycoprotein (G) was developed and laboratory evaluated using sera from experimental pigs, mini pigs and nonhuman primates. The test depends on competition between specific antibodies in positive sera and a virus–specific mAb for binding to NiV–G. Results: Based on 1,199 negative and 71 NiV positive serum test results, the cutoff value was determined as 35% inhibition. The diagnostic sensitivity and specificity of the NiV cELISA was 98.58 and 99.92%, respectively. When testing sera from animals experimentally infected with NiV Malaysia, the cELISA detected antibodies from 14 days post–infection (dpi) and remained positive until the end of the experiment (28 dpi). Comparisons using the Kappa coefficient showed strong agreement (100%) between the cELISA and a plaque reduction neutralization test (PRNT). Discussion: Because our cELISA is simpler, faster, and gives comparable or better results than PRNT, it would be an adequate screening test for suspect NiV and HeV cases, and it would also be useful for epidemiological surveillance of Henipavirus infections in different animal species without changing reagents. KEYWORDS Nipah virus, Hendra virus, Henipavirus, recombinant Nipah virus glycoprotein, monoclonal antibody, competitive ELISA, serodiagnosis Introduction Nipah virus (NiV) and Hendra virus (HeV) are enveloped, negative–sense RNA viruses that belong to the genus Henipavirus in the virus family Paramyxoviridae. These zoonotic viruses cause fatal diseases in animals and humans (1–3). NiV was first observed in Malaysia and Singaporean pigs in 1998–99, with 105 confirmed human deaths. Since then, NiV infections have been identified in both animals and humans in South and Southeast Asia (1, 4, 5). HeV was first identified during a disease outbreak in 1994 in Australia, where several horses and their Frontiers in Veterinary Science 01 frontiersin.org