title: Identification of a major immunogenic region of equine herpesvirus-1 glycoprotein E and its application to enzyme-linked immunosorbent assay.
abstract: A major immunogenic region of equine herpesvirus (EHV)-1 glycoprotein E (gE) was identified. Firstly, the various fragments of EHV-1 gE were expressed as fusion proteins with glutathione S-transferase (GST) in Escherichia coli and their antigenicities were compared by immunoblot analysis using sera from horses experimentally infected with EHV-1. Thirty-three amino acids of gE (a.a. 169-201) specifically and sensitively reacted with the antibodies induced by EHV-1 but not EHV-4 infection. The corresponding region of EHV-4 gE (a.a. 169-199) did not react with antibodies to EHV-1, indicating that this region is specific for each virus. In addition, when the antigenicities of three 20-mer synthetic peptides of EHV-1 gE, gE1(169-188), gE1(176-195) and gE1(182-201) were compared by enzyme-linked immunosorbent assay (ELISA), gE1(169-188) was found to contain a major B-cell epitope. ELISA using two synthetic peptides, gE1(169-188) and gG4(319-330), previously identified as the major EHV-4-specific epitope in gG, was developed and could specifically detect antibodies to EHV-1 and EHV-4, respectively. In Japan, the EHV-1 deleted in gE gene (EHV-1 ΔgE) virus is expected to be introduced in the field as a new modified live vaccine. This ELISA did not react with antibodies induced by inoculation with EHV-1 ΔgE, indicating that it is a useful method to differentiate between EHV-1 infection and EHV-1 ΔgE inoculation. In conclusion, the ELISA described herein, using synthetic peptides, is a simple method to distinguish between EHV-1 and EHV-4 infections and will be suitable as a vaccine marker after introduction of EHV-1 ΔgE into field horses.
authors: Laboratory of Veterinary Microbiology, Joint Faculty of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors: Kiyohiko
authors: K
authors: Andoh
authors: Maaya
authors: M
authors: Takasugi
authors: Hassan Y A H
authors: HY
authors: Mahmoud
authors: Shiho
authors: S
authors: Hattori
authors: Yutaka
authors: Y
authors: Terada
authors: Keita
authors: K
authors: Noguchi
authors: Hiroshi
authors: H
authors: Shimoda
authors: Hiroshi
authors: H
authors: Bannai
authors: Koji
authors: K
authors: Tsujimura
authors: Tomio
authors: T
authors: Matsumura
authors: Takashi
authors: T
authors: Kondo
authors: Ken
authors: K
authors: Maeda
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D018985:Epitopes, B-Lymphocyte / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D004861:Herpesvirus 1, Equid; D023141:Herpesvirus 4, Equid; D006734:Horse Diseases / Q000175:diagnosis*; D006736:Horses; D014759:Viral Envelope Proteins / Q000302:isolation & purification*
chemical_list: D018985:Epitopes, B-Lymphocyte; C111143:Equid herpesvirus 1 gE; D014759:Viral Envelope Proteins