A type-specific serological test to distinguish antibodies to equine herpesviruses 4 and 1.
- Comparative Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
This research introduces a unique testing method known as Enzyme-Linked Immunosorbent Assay (ELISA), which identifies horses that have been infected with either Equine Herpesvirus 4 (EHV4) or EHV1. The test uses specific antigens associated with these viruses and proves to be highly effective in differentiating between the two infections.
Enzyme-Linked Immunosorbent Assay (ELISA)
This study illustrates the development of a type-specific ELISA. This tool significantly helps in distinguishing antibodies to equine herpesviruses 4 (EHV4) and 1 (EHV1).
- ELISA allows researchers to identify if horses have been infected with either of these antigenically similar viruses.
- The specific antigens used for the ELISA are parts of EHV4 and EHV1 glycoprotein G (gG) homologues.
- These are expressed in E. coli as fusion proteins, giving them the ability to facilitate the differentiating process.
Studied Horses and Test Results
In the study, a mix of Thoroughbred and Standardbred horses were involved.
- The researchers ran tests on 97 Thoroughbred and 174 Standardbred horses, and all were found to be strongly EHV4 ELISA positive while 30% were EHV1 ELISA positive.
- These horses were chosen wisely, considering they were all unvaccinated, which removes the influence of any vaccine-induced antibodies.
Relevance and Effectiveness of the Test
The researchers also examined the specificity and sensitivity of the ELISA tool.
- The type-specificity of the EHV1 gG antigen was tested in cross-absorption experiments.
- 96% of EHV1 ELISA positive horses were found to be genuine EHV1 antibody positives, highlighting the reliability of the ELISA tool.
- Maintenance of EHV1 gG antibody was examined by testing the sera obtained from mares four years after a confirmed EHV1 abortion. Seven out of 10 mares remained EHV1 ELISA positive.
- This information reflects the persistency of these antibodies over time, crucial for understanding the pathogenesis and long-term immune response to the EHV1.
In conclusion, the ELISA tool devised in this research is precise and sensitive enough to detect all horses previously infected with EHV4, and most previously infected with EHV1. It has the potential to substantially improve EHV disease management and prevention strategies.
Cite This Article
Publication
Researcher Affiliations
- Centre for Equine Virology, School of Veterinary Science, University of Melbourne, Parkville, Victoria, Australia.
MeSH Terms
- Animals
- Antibodies, Viral / blood
- Base Sequence
- DNA, Viral / blood
- Enzyme-Linked Immunosorbent Assay / methods
- Enzyme-Linked Immunosorbent Assay / veterinary
- Female
- Herpesviridae Infections / diagnosis
- Herpesviridae Infections / veterinary
- Herpesvirus 1, Equid / immunology
- Herpesvirus 1, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horses
- Leukocytes / virology
- Molecular Sequence Data
- Polymerase Chain Reaction
- Recombinant Fusion Proteins / immunology
- Sensitivity and Specificity
- Viral Envelope Proteins / immunology
References
- Smith DB, Johnson KS. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase.. Gene 1988 Jul 15;67(1):31-40.
- Telford EA, Watson MS, McBride K, Davison AJ. The DNA sequence of equine herpesvirus-1.. Virology 1992 Jul;189(1):304-16.
- Colle CF 3rd, Flowers CC, O'Callaghan DJ. Open reading frames encoding a protein kinase, homolog of glycoprotein gX of pseudorabies virus, and a novel glycoprotein map within the unique short segment of equine herpesvirus type 1.. Virology 1992 Jun;188(2):545-57.
- Fitzpatrick DR, Studdert MJ. Immunologic relationships between equine herpesvirus type 1 (equine abortion virus) and type 4 (equine rhinopneumonitis virus).. Am J Vet Res 1984 Oct;45(10):1947-52.
- Allen GP, Bryans JT. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections.. Prog Vet Microbiol Immunol 1986;2:78-144.
- Drummer HE, Reynolds A, Studdert MJ, MacPherson CM, Crabb BS. Application of an equine herpesvirus 1 (EHV1) type-specific ELISA to the management of an outbreak of EHV1 abortion.. Vet Rec 1995 Jun 10;136(23):579-81.
- Peet RL, Coackley W, Smith VW, Main C. Equine abortion associated with herpesvirus.. Aust Vet J 1978 Mar;54(3):151.
- Studdert MJ, Crabb BS, Ficorilli N. The molecular epidemiology of equine herpesvirus 1 (equine abortion virus) in Australasia 1975 to 1989.. Aust Vet J 1992 May;69(5):104-11.
- Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.. Science 1988 Jan 29;239(4839):487-91.
- Crabb BS, Nagesha HS, Studdert MJ. Identification of equine herpesvirus 4 glycoprotein G: a type-specific, secreted glycoprotein.. Virology 1992 Sep;190(1):143-54.
- Crabb BS, Studdert MJ. Equine herpesviruses 4 (equine rhinopneumonitis virus) and 1 (equine abortion virus).. Adv Virus Res 1995;45:153-90.
- Westbury HA, Kovesdy L, Barton MD. Natural intra-uterine infection of foals with equine herpes virus type 1 in Victoria.. Aust Vet J 1978 Mar;54(3):147.
- Crabb BS, Studdert MJ. Comparative studies of the proteins of equine herpesviruses 4 and 1 and asinine herpesvirus 3: antibody response of the natural hosts.. J Gen Virol 1990 Sep;71 ( Pt 9):2033-41.
- Welch HM, Bridges CG, Lyon AM, Griffiths L, Edington N. Latent equid herpesviruses 1 and 4: detection and distinction using the polymerase chain reaction and co-cultivation from lymphoid tissues.. J Gen Virol 1992 Feb;73 ( Pt 2):261-8.
- Slater JD, Borchers K, Thackray AM, Field HJ. The trigeminal ganglion is a location for equine herpesvirus 1 latency and reactivation in the horse.. J Gen Virol 1994 Aug;75 ( Pt 8):2007-16.
- Dixon RJ, Hartley WJ, Hutchins DR, Lepherd EE, Feilen C, Jones RF, Love DN, Sabine M, Wells AL. Perinatal foal mortality associated with a herpesvirus.. Aust Vet J 1978 Mar;54(3):103-5.
- Crabb BS, Allen GP, Studdert MJ. Characterization of the major glycoproteins of equine herpesviruses 4 and 1 and asinine herpesvirus 3 using monoclonal antibodies.. J Gen Virol 1991 Sep;72 ( Pt 9):2075-82.
- Allen GP, Yeargan MR, Turtinen LW, Bryans JT, McCollum WH. Molecular epizootiologic studies of equine herpesvirus-1 infections by restriction endonuclease fingerprinting of viral DNA.. Am J Vet Res 1983 Feb;44(2):263-71.
- Studdert MJ, Blackney MH. Equine herpesviruses: on the differentiation of respiratory from foetal strains of type 1.. Aust Vet J 1979 Oct;55(10):488-92.
- Crabb BS, Studdert MJ. Epitopes of glycoprotein G of equine herpesviruses 4 and 1 located near the C termini elicit type-specific antibody responses in the natural host.. J Virol 1993 Oct;67(10):6332-8.
Citations
This article has been cited 13 times.- El Brini Z, Fassi Fihri O, Paillot R, Lotfi C, Amraoui F, El Ouadi H, Dehhaoui M, Colitti B, Alyakine H, Piro M. Seroprevalence of Equine Herpesvirus 1 (EHV-1) and Equine Herpesvirus 4 (EHV-4) in the Northern Moroccan Horse Populations.. Animals (Basel) 2021 Sep 29;11(10).
- Scheurer L, Bachofen C, Hardmeier I, Lechmann J, Schoster A. Prevalence of Nasal Shedding of Equid Gammaherpesviruses in Healthy Swiss Horses.. Viruses 2021 Aug 25;13(9).
- Attili AR, Colognato R, Preziuso S, Moriconi M, Valentini S, Petrini S, De Mia GM, Cuteri V. Evaluation of Three Different Vaccination Protocols against EHV1/EHV4 Infection in Mares: Double Blind, Randomized Clinical Trial.. Vaccines (Basel) 2020 Jun 1;8(2).
- Azab W, Bedair S, Abdelgawad A, Eschke K, Farag GK, Abdel-Raheim A, Greenwood AD, Osterrieder N, Ali AAH. Detection of equid herpesviruses among different Arabian horse populations in Egypt.. Vet Med Sci 2019 Aug;5(3):361-371.
- Muscat KE, Padalino B, Hartley CA, Ficorilli N, Celi P, Knight P, Raidal S, Gilkerson JR, Muscatello G. Equine Transport and Changes in Equid Herpesvirus' Status.. Front Vet Sci 2018;5:224.
- Maxwell LK, Bentz BG, Gilliam LL, Ritchey JW, Pusterla N, Eberle R, Holbrook TC, McFarlane D, Rezabek GB, Meinkoth J, Whitfield C, Goad CL, Allen GP. Efficacy of the early administration of valacyclovir hydrochloride for the treatment of neuropathogenic equine herpesvirus type-1 infection in horses.. Am J Vet Res 2017 Oct;78(10):1126-1139.
- Björnsdóttir S, Harris SR, Svansson V, Gunnarsson E, Sigurðardóttir ÓG, Gammeljord K, Steward KF, Newton JR, Robinson C, Charbonneau ARL, Parkhill J, Holden MTG, Waller AS. Genomic Dissection of an Icelandic Epidemic of Respiratory Disease in Horses and Associated Zoonotic Cases.. mBio 2017 Aug 1;8(4).
- Mahmoud HY, Andoh K, Hattori S, Terada Y, Noguchi K, Shimoda H, Maeda K. Characterization of glycoproteins in equine herpesvirus-1.. J Vet Med Sci 2013 Oct;75(10):1317-21.
- Smith KL, Li Y, Breheny P, Cook RF, Henney PJ, Sells S, Pronost S, Lu Z, Crossley BM, Timoney PJ, Balasuriya UB. New real-time PCR assay using allelic discrimination for detection and differentiation of equine herpesvirus-1 strains with A2254 and G2254 polymorphisms.. J Clin Microbiol 2012 Jun;50(6):1981-8.
- Van de Walle GR, Sakamoto K, Osterrieder N. CCL3 and viral chemokine-binding protein gg modulate pulmonary inflammation and virus replication during equine herpesvirus 1 infection.. J Virol 2008 Feb;82(4):1714-22.
- Maeda K, Mizukoshi F, Hamano M, Kai K, Kondo T, Matsumura T. Identification of another B-cell epitope in the type-specific region of equine herpesvirus 4 glycoprotein G.. Clin Diagn Lab Immunol 2005 Jan;12(1):122-4.
- Maeda K, Mizukoshi F, Hamano M, Kai K, Iwata H, Kondo T, Matsumura T. Development of an equine herpesvirus type 4-specific enzyme-linked immunosorbent assay using a B-cell epitope as an antigen.. J Clin Microbiol 2004 Mar;42(3):1095-8.
- Duffy MF, Whithear KG, Noormohammadi AH, Markham PF, Catton M, Leydon J, Browning GF. Indirect enzyme-linked immunosorbent assay for detection of immunoglobulin G reactive with a recombinant protein expressed from the gene encoding the 116-kilodalton protein of Mycoplasma pneumoniae.. J Clin Microbiol 1999 Apr;37(4):1024-9.