title: 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.
authors.affiliation: Epizootic Research Center, Equine Research Institute, Japan Racing Association, 1400-4 Shiba, Shimotsuke, Tochigi 329-0412, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Bannai
authors.forename: Manabu
authors.initials: M
authors.lastname: Nemoto
authors.forename: Koji
authors.initials: K
authors.lastname: Tsujimura
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamanaka
authors.forename: Takashi
authors.initials: T
authors.lastname: Kondo
authors.forename: Tomio
authors.initials: T
authors.lastname: Matsumura
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing; D002417:Cattle; D002460:Cell Line; D002482:Cellulose; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D004861:Herpesvirus 1, Equid / Q000276:immunology*; D057166:High-Throughput Screening Assays / Q000662:veterinary; D006734:Horse Diseases / Q000175:diagnosis* / Q000821:virology*; D006736:Horses; D009500:Neutralization Tests / Q000379:methods / Q000662:veterinary*
chemical_list: D057134:Antibodies, Neutralizing; D002482:Cellulose