Identification of equine herpesvirus 3 (equine coital exanthema virus), equine gammaherpesviruses 2 and 5, equine adenoviruses 1 and 2, equine arteritis virus and equine rhinitis A virus by polymerase chain reaction.
Abstract: To develop rapid (< 8 hour) tests using polymerase chain reaction (PCR) for the diagnosis of equine herpesvirus 3 (EHV3; equine coital exanthema virus), equine gammaherpesviruses 2 (EHV2) and EHV5, equine adenovirus 1 (EAdV1), EAdV2, equine arteritis virus (EAV), equine rhinitis A virus (ERAV; formerly equine rhinovirus 1) Methods: Either single round or second round (seminested) PCRs were developed and validated. Methods: Oligonucleotide primers were designed that were specific for each virus, PCR conditions were defined and the specificity and sensitivity of the assays were determined. The application of the tests was validated using a number of independent virus isolates for most of the viruses studied. The PCRs were applied directly to clinical samples where samples were available. Results: We developed a single round PCR for the diagnosis of EHV3, a seminested PCR for EHV2 and single round PCRs for EHV5, EAdV1, EAdV2 and RT-PCRs for EAV and ERAV. The PCR primer sets for each virus were designed and shown to be highly specific (did not amplify any recognised non-target template) and sensitive (detection of minimal amounts of virus) and, where multiple virus isolates were available all isolates were detected. Conclusions: The development and validation of a comprehensive panel of PCR diagnostic tests, predominantly for viruses causing equine respiratory disease, that can be completed within 8 hours from receipt of clinical samples, provides a major advance in the rapid diagnosis or exclusion diagnosis of these endemic equine virus diseases in Australia.
Publication Date: 2001-11-20 PubMed ID: 11712710DOI: 10.1111/j.1751-0813.2001.tb10674.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Clinical Study
- Diagnosis
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Management
- Disease Outbreaks
- Epidemiology
- Equine Diseases
- Equine Health
- Equine Herpesvirus
- Equine Rhinitis Virus
- Equine Viral Arteritis
- Infectious Disease
- Laboratory Methods
- Polymerase Chain Reaction
- Veterinary Care
- Veterinary Medicine
- Veterinary Research
- Virology
- Virus
Summary
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The research revolves around the development of rapid diagnostic tests using the Polymerase Chain Reaction (PCR) methodology for several equine viruses. These efficient tests helped identify specific types of equine viruses within 8 hours of receiving clinical samples.
Methodology
- The study involved creating new tests utilizing the Polymerase Chain Reaction method for the identification of various equine viruses including Equine Herpes Virus (EHV3, EHV2, EHV5), Equine Adenovirus 1(EAdV1), EAdV2, Equine Arteritis Virus (EAV) and Equine rhinitis A virus (ERAV).
- Specific oligonucleotide primers for each virus type were carefully designed to ensure accurate results.
- Depending on the type of virus being identified, either single round or second round (seminested) PCRs were developed and used.
- Once the PCR conditions were defined, the specificity (ability not to respond to non-target entities) and sensitivity (ability to detect minimal amounts of virus) of these tests were determined.
- The researchers further validated the application of these tests using multiple independent virus isolates for most of the viruses being studied. Nor wherever possible, these PCRs were applied directly to clinical samples.
Results
- The researchers successfully created a single round PCR for EHV3 diagnosis, a seminested PCR for EHV2, single round PCRs for EHV5, EAdV1, EAdV2, and RT-PCRs for EAV and ERAV.
- The PCR primer sets designed for each virus were found to be highly specific and sensitive, capable of detecting even minimal amounts of each virus type.
- Every virus isolate available for the study was detected using the newly created tests, providing further validation of their effectiveness.
Conclusions
- The study led to the development and confirmation of a comprehensive panel of PCR diagnostic tests, mainly for equine respiratory diseases causing viruses.
- The significant feature of these tests is their rapid detection time, with the ability to provide results within 8 hours of receiving clinical samples.
- This novel approach can significantly improve the rapid diagnosis or exclusion diagnosis of these prevalent equine virus diseases, particularly in Australia.
Cite This Article
APA
Dynon K, Varrasso A, Ficorilli N, Holloway S, Reubel G, Li F, Hartley C, Studdert M, Drummer H.
(2001).
Identification of equine herpesvirus 3 (equine coital exanthema virus), equine gammaherpesviruses 2 and 5, equine adenoviruses 1 and 2, equine arteritis virus and equine rhinitis A virus by polymerase chain reaction.
Aust Vet J, 79(10), 695-702.
https://doi.org/10.1111/j.1751-0813.2001.tb10674.x Publication
Researcher Affiliations
- Centre for Equine Virology, School of Veterinary Science, The University of Melbourne, Parkville, Victoria.
MeSH Terms
- Adenoviridae / genetics
- Adenoviridae / isolation & purification
- Animals
- Aphthovirus / classification
- Aphthovirus / genetics
- Aphthovirus / isolation & purification
- Base Sequence
- DNA Primers
- Equartevirus / classification
- Equartevirus / genetics
- Equartevirus / isolation & purification
- Herpesvirus 3, Equid / classification
- Herpesvirus 3, Equid / genetics
- Herpesvirus 3, Equid / isolation & purification
- Horse Diseases / diagnosis
- Horse Diseases / virology
- Horses
- Polymerase Chain Reaction / veterinary
- Respiratory Tract Infections / diagnosis
- Respiratory Tract Infections / veterinary
- Respiratory Tract Infections / virology
- Rhadinovirus / classification
- Rhadinovirus / genetics
- Rhadinovirus / isolation & purification
- Sensitivity and Specificity
- Virus Diseases / diagnosis
- Virus Diseases / veterinary
- Virus Diseases / virology
- Viruses / classification
- Viruses / genetics
- Viruses / isolation & purification
Citations
This article has been cited 16 times.- Troncoso I, Calvanese R, Saravia F, Muñoz-Leal S, Zegpi NA, Ortega R. First molecular detection of Equine Herpesvirus type 3 (EHV-3) in Chile. Vet Med Sci 2023 Mar;9(2):717-720.
- Onasanya AE, El-Hage C, Diaz-Méndez A, Vaz PK, Legione AR, Browning GF, Devlin JM, Hartley CA. Whole genome sequence analysis of equid gammaherpesvirus -2 field isolates reveals high levels of genomic diversity and recombination. BMC Genomics 2022 Aug 30;23(1):622.
- Lee SK, Choi J, Yoon J, Jung J, Park JY, Park J, Kim Y, Park JY, Park D. Molecular Detection of Equine Adenovirus 1 in Nasal Swabs from Horses in the Republic of Korea. Vet Sci 2022 Apr 13;9(4).
- El-Hage C, Mekuria Z, Dynon K, Hartley C, McBride K, Gilkerson J. Association of Equine Herpesvirus 5 with Mild Respiratory Disease in a Survey of EHV1, -2, -4 and -5 in 407 Australian Horses. Animals (Basel) 2021 Nov 30;11(12).
- Uchida-Fujii E, Kinoshita Y, Niwa H, Maeda T, Nukada T, Ueno T. High prevalence of Mycoplasma equirhinis in Thoroughbred horses with respiratory symptoms in autumn 2018. J Vet Med Sci 2021 Dec 9;83(12):1907-1912.
- Anis E, Ilha MRS, Engiles JB, Wilkes RP. Evaluation of targeted next-generation sequencing for detection of equine pathogens in clinical samples. J Vet Diagn Invest 2021 Mar;33(2):227-234.
- Toishi Y, Tsunoda N, Kirisawa R. Period of excretion of equine herpesvirus 3 (EHV-3) from a stallion before showing clinical signs of equine coital exanthema and the effect of acyclovir treatment on the duration of EHV-3 excretion. J Vet Med Sci 2020 Sep 24;82(9):1299-1305.
- 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.
- Seeber PA, Dayaram A, Sicks F, Osterrieder N, Franz M, Greenwood AD. Noninvasive Detection of Equid Herpesviruses in Fecal Samples. Appl Environ Microbiol 2019 Feb 1;85(3).
- 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.
- 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).
- Kirisawa R, Toishi Y, Akamatsu A, Soejima K, Miyashita T, Tsunoda N. Isolation of equine herpesvirus 3 (EHV-3) from equine coital exanthema of two stallions and sero-epidemiology of EHV-3 infection in Japan. J Vet Med Sci 2017 Mar 23;79(3):636-643.
- Williams KJ, Robinson NE, Lim A, Brandenberger C, Maes R, Behan A, Bolin SR. Experimental induction of pulmonary fibrosis in horses with the gammaherpesvirus equine herpesvirus 5. PLoS One 2013;8(10):e77754.
- Marenzoni ML, Bietta A, Lepri E, Casagrande Proietti P, Cordioli P, Canelli E, Stefanetti V, Coletti M, Timoney PJ, Passamonti F. Role of equine herpesviruses as co-infecting agents in cases of abortion, placental disease and neonatal foal mortality. Vet Res Commun 2013 Dec;37(4):311-7.
- Lu Z, Timoney PJ, White J, Balasuriya UB. Development of one-step TaqMan® real-time reverse transcription-PCR and conventional reverse transcription-PCR assays for the detection of equine rhinitis A and B viruses. BMC Vet Res 2012 Jul 25;8:120.
- Marenzoni ML, Coppola G, Maranesi M, Passamonti F, Cappelli K, Capomaccio S, Verini Supplizi A, Thiry E, Coletti M. Age-dependent prevalence of equid herpesvirus 5 infection. Vet Res Commun 2010 Dec;34(8):703-8.
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