AccScience Publishing / MI / Online First / DOI: 10.36922/MI025470126
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ORIGINAL RESEARCH ARTICLE

Evaluation of chicken egg yolk immunoglobulin Y-based ELISA for serodiagnosis of hepatitis E virus in swine

Arnaud Fondjo Kouam1,2* ,  Philipe Herman Nfombouot Njitoyap2 ,  Brice Fredy Nemg Simo3 ,  Cromwel Tepap Zemnou2 ,  Kerinyuy Juliene Kongnyuy1 ,  Armel Jackson Seukep1 ,  Elisabeth Menkem Zeuko’o1 ,  Frédéric Nico Njayou2 ,  Paul Fewou Moundipa2
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1 Department of Biomedical Sciences, Faculty of Health Sciences, University of Buea, Buea, Southwest Region, Cameroon
2 Department of Biochemistry, Faculty of Science, University of Yaoundé 1, Yaoundé, Centre Region, Cameroon
3 The Centre for Research in Infectious Diseases, Yaoundé, Centre Region, Cameroon
MI, 025470126 https://doi.org/10.36922/MI025470126
Received: 21 November 2025 | Revised: 28 December 2025 | Accepted: 26 January 2026 | Published online: 19 February 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Hepatitis E virus (HEV) is a major zoonotic pathogen, and pigs are an important reservoir. Serology for anti-HEV antibodies poorly reflects active infection, while reverse transcription polymerase chain reaction (RT-PCR) is sensitive but not always feasible for large-scale surveillance. This study aimed to develop an immunoglobulin Y (IgY)-based sandwich enzyme-linked immunosorbent assay (ELISA) for direct detection of HEV antigen (HEV-Ag). An antigenic epitope from the HEV open reading frame 2 capsid protein was identified in silico, synthesized, and used to immunize laying hens; IgY was purified from egg yolk and characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis. An in-house sandwich ELISA was subsequently developed using the anti-HEV IgY antibodies. Its diagnostic performance was evaluated against RT-PCR using 350 field-collected pig serum samples and compared with commercial anti-HEV IgG and IgM ELISA kits. Immunization yielded high-purity IgY antibodies, with antigen-specific antibody kinetics showing a significant increase in specific immunoreactivity from week 4 onward and peak sensitivity at weeks 6–8 post-immunization. The anti-HEV IgY showed high specificity, with no cross-reactivity observed against a heterologous antigen (HBsAg). The optimized sandwich ELISA detected HEV-Ag with a sensitivity of 95.4% and a specificity of 100% relative to RT-PCR. The prevalence of HEV-Ag was 17.7% (62/350), which was not significantly different from RT-PCR (18.6%, 65/350; p=0.250). In contrast, commercial IgG and IgM ELISAs showed high seroprevalence rates (34.9% and 28.3%, respectively) but exhibited significant discrepancies with RT-PCR (p<0.0001), indicating limited utility for identifying active infection. Overall, the developed IgY-based sandwich ELISA demonstrated high diagnostic performance for detecting active HEV infection. This assay represents a viable alternative to RT-PCR for large-scale surveillance, particularly in resource-constrained settings, and may contribute to improved control of zoonotic HEV transmission.

Graphical abstract
Keywords
Hepatitis E virus
IgY antibodies
Sandwich ELISA
Antigen detection
Pig serum
Diagnostic performance
Funding
This research was supported by the International Center for Genetic Engineering and Biotechnology (ICGEB) (grant number: S/CMR15-06), and Ministry of Higher Education of Cameroon under Trimester Research Modernization Allocation.
Conflict of interest
The authors declare that they have no competing interests.
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Microbes & Immunity, Electronic ISSN: 3029-2883 Print ISSN: 3041-0886, Published by AccScience Publishing
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