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
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- Journal Article
Summary
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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 5 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.
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