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Development of a focus-reduction neutralizing test for detecting equine herpesvirus type-1-neutralizing antibodies.
Abstract: Virus-neutralizing (VN) testing is essential for evaluating virus-specific immunity in equine herpesvirus type-1 (EHV-1) infection. We developed a focus-reduction neutralization test (FRNT) for EHV-1 using 96-well plates for faster large-scale testing with sufficient sensitivity. We used an overlay medium containing Avicel (FMC Biopolymer), a microcrystalline cellulose with lower viscosity than the methylcellulose. The foci were visualized by immuno-staining with anti-EHV-1 gp14 monoclonal antibody. The FRNT successfully detected seroconversion in horses experimentally infected with EHV-1 (n = 3) and in those infected naturally (n = 16). The FRNT for EHV-1 was high-throughput and time saving. The FRNT can be used in large-scale seroepidemiological studies of EHV-1 and in evaluating vaccine efficacy.
Publication Date: 2013-04-16 PubMed ID: 23595119DOI: 10.1292/jvms.13-0043Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study presents the development of a faster and more sensitive method for testing immunity to equine herpesvirus type-1 (EHV-1) in horses, both naturally and experimentally infected. The method, named focus-reduction neutralization test (FRNT), employs a microcrystalline cellulose medium and an anti-EHV-1 gp14 monoclonal antibody for accurate detection of the virus.
Context and Importance of Research
- This research is primarily concerned with the improved detection of equine herpesvirus type-1 (EHV-1), a common virus in horses that can cause respiratory disease, abortion, neonatal death, and neurological disorders.
- Effective virus-neutralizing (VN) testing is critical in assessing virus-specific immunity, understanding the spread of viruses in populations, and verifying the effectiveness of vaccines.
Development of the FRNT Method
- The researchers introduced a new focus-reduction neutralization test (FRNT) for EHV-1 in this study.
- This novel approach uses 96-well plates, which make the process faster and more efficient for large-scale testing.
- For better visualization of the virus, the foci (areas of infection) were made visible by immuno-staining with an anti-EHV-1 gp14 monoclonal antibody.
- Instead of the typical methylcellulose, Avicel—a microcrystalline cellulose with a lower viscosity—was used in the overlay medium.
Implementation and Results of the New Test
- To verify its effectiveness, the FRNT method was used to detect seroconversion in horses that were either naturally infected (16 horses) or experimentally infected (3 horses) with EHV-1.
- The results revealed that the FRNT for EHV-1 was high-throughput, saving time and proving suitable for use in large-scale seroepidemiological studies.
Implications and Future Use of FRNT
- The successful results of this study indicate that the FRNT method would be valuable for gathering larger-scale epidemiological data on EHV-1.
- Furthermore, the FRNT could be used to evaluate the efficacy of EHV-1 vaccines, thus aiding in the development and assessment of future treatments.
Cite This Article
APA
Bannai H, Nemoto M, Tsujimura K, Yamanaka T, Kondo T, Matsumura T.
(2013).
Development of a focus-reduction neutralizing test for detecting equine herpesvirus type-1-neutralizing antibodies.
J Vet Med Sci, 75(9), 1209-1212.
https://doi.org/10.1292/jvms.13-0043 Publication
Researcher Affiliations
- Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
MeSH Terms
- Animals
- Antibodies, Neutralizing
- Cattle
- Cell Line
- Cellulose
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / immunology
- High-Throughput Screening Assays / veterinary
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Neutralization Tests / methods
- Neutralization Tests / veterinary
Citations
This article has been cited 7 times.- Vaidya SR. Immuno-Colorimetric Neutralization Test: A Surrogate for Widely Used Plaque Reduction Neutralization Tests in Public Health Virology. Viruses 2023 Apr 10;15(4).
- Bannai H, Kambayashi Y, Tsujimura K, Nagashima T, Takebe N, Tominari M, Nemoto M, Ohta M. Persistence of virus-neutralizing antibodies in horses inoculated with two doses of a live equine herpesvirus type 1 vaccine with different vaccination intervals. J Equine Sci 2021;32(3):99-102.
- Pavulraj S, Eschke K, Theisen J, Westhoff S, Reimers G, Andreotti S, Osterrieder N, Azab W. Equine Herpesvirus Type 4 (EHV-4) Outbreak in Germany: Virological, Serological, and Molecular Investigations. Pathogens 2021 Jun 25;10(7).
- Bannai H, Tsujimura K, Nemoto M, Ohta M, Yamanaka T, Kokado H, Matsumura T. Epizootiological investigation of equine herpesvirus type 1 infection among Japanese racehorses before and after the replacement of an inactivated vaccine with a modified live vaccine. BMC Vet Res 2019 Aug 6;15(1):280.
- Bannai H, Mae N, Ode H, Nemoto M, Tsujimura K, Yamanaka T, Kondo T, Matsumura T. Successful control of winter pyrexias caused by equine herpesvirus type 1 in Japanese training centers by achieving high vaccination coverage. Clin Vaccine Immunol 2014 Aug;21(8):1070-6.
- Bannai H, Kambayashi Y, Kume K, Takebe N, Endo Y, Kawanishi N, Nemoto M, Tsujimura K. Reduction in endemic equine herpesvirus type-1 and type-4 infection among Thoroughbred yearlings through an updated vaccination program. J Equine Sci 2025 Jun;36(2):67-74.
- El Brini Z, Cullinane A, Garvey M, Fassi Fihri O, Fellahi S, Amraoui F, Loutfi C, Sebbar G, Paillot R, Piro M. First Molecular and Phylogenetic Characterization of Equine Herpesvirus-1 (EHV-1) and Equine Herpesvirus-4 (EHV-4) in Morocco. Animals (Basel) 2025 Jan 5;15(1).
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