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Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction

Year 2023, Volume: 6 Issue: 4, 705 - 712, 15.10.2023
https://doi.org/10.19127/bshealthscience.1349444

Abstract

The aim of this study was to use polymerase chain reaction (PCR) to detect and differentiate Equine Herpesvirus Type-1 (EHV-1) and Type-4 (EHV-4) viruses within the racehorse population in Türkiye. The diagnostic sensitivity of PCR was also assessed. For this purpose, 98 nasal swab specimens from naturally infected racehorses aged 2 and above, displaying respiratory symptoms suggestive of EHV infections, and 26 aborted fetuses with various organ samples were collected. DNA extraction and PCR analysis were performed on these samples. The study started with PCR optimization using reference strains of EHV-1 (89c25p) and EHV-4 (TH20p) DNA. Finally optimized was completed and reference strains were used for this study. In conclusion, PCR successfully detected and differentiated 7 EHV-1 positive samples from the tissues of the 26 aborted fetuses, as well as one EHV-1 positive and two EHV-4 positive samples from the nasal swabs of the 98 cases. This study represents one of the pioneering works where PCR was firstly employed to detect and differentiate EHV-1 and EHV-4 strains in Türkiye. The study's findings reveal the presence of both EHV-1 and EHV-4 in Türkiye's racehorse population and is among the early reports to identify the existence of EHV-4 using PCR. These findings underscore the circulation of both viruses within the racehorse population. As a result of this study, it has been concluded that the PCR method is a sensitive, cost-effective, and time-saving diagnostic approach for detecting and distinguishing EHV-1 and EHV-4 infections.

Project Number

Doktora-VFK231T

References

  • Allen GP, Bryans JT. 1986. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections. Prog Vet Microbiol Immun, 2: 78-144.
  • Ataseven VS, Bilge-Dağalp S, Başaran Z, Keskin S 2010. Seroepidemiological studies of equine herpesviruses 1 (EHV-1) and 4 (EHV-4) infections in working horses from the eastern Turkey. Ankara Üniv Vet Fak Derg, 57: 39-42.
  • Ballagi-Pordany A, Klingeborn B, Flensburg J, Belak S. 1990. Equine herpesvirus type1: Detection of viral DNA sequences in aborted fetuses with the polymerase chain reaction. Vet Microbiol, 22: 373-381.
  • Başkaya H, Keskintepe H, Doğuer M, İyigören B, Yılmaz S, Demir A. 1968. An outbreak of equine virus abortion in Turkey. Isolation and identification of rhino pneumonitis virus in cell cultures. A Ü Vet Fak Derg, 15 (3-4): 309-316.
  • Carvalho R, Passos LMF, Martins AS. 2000. Development of a differential multiplex PCR assay for equine herpesvirus 1 and 4 as a diagnostic tool. J Vet Med, 47:351-359.
  • Carvalho R, Oliveira AM, Souza AM, Passos LMF, Martins AS. 2000. Prevalence of equine herpesvirus type 1 latency detected by polymerase chain reaction. Arch Virol, 145: 1773-1787.
  • Crabb BS, Studdert MJ. 1993. 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, 10: 6332-6338.
  • Csellner H, Walker C, Wellington JE, McLure LE, Love DN, Whalley JM. 2000. EHV-1glycoprotein D (EHV-1 gD) is required for virus entry and cell-cell fusion, and an EHV-1 gD deletion mutant induces a protective immune response in mice. Arch Virol, 145: 2371-2385.
  • Damiani AM, Matsumura T, Yokoyama N, Takeshi M, Takahashi E. 2000. A deletion in the gI and gE genes of equine herpesvirus type 4 reduces viral virulence in the natural host and affects virus transmission during cell to cell spread. Virus Res, 67: 189-202.
  • Damiani AM, Matsumura T, Jang HK, Izumiya Y, Milkami T, Takahashi E. 2000. Identification of the products of the equine herpesvirus type 4 gI and gE genes. Arch Virol, 145: 1489-1496.
  • Donaldson MT, Sweeney CR. 1998. Herpesvirus myeloencephalopathy in horses: 11 cases (1982-1996). J Amer Vet Med Assoc, 213 (5): 671-675.
  • Drummer HE, Reynolds A, Studdert MJ, MacPherson CM, Crabb BS. 1995. Application of an equine herpesvirus 1 (EHV-1) type-specific ELISA to the management of an outbreak of EHV1 abortion. Vet Rec, 136: 579-581.
  • Edington N, Welch HM, Griffiths L. 1994. The prevalence of latent equid herpesviruses in the tissues of 40 abattoir horses. Equine Vet J, 26(2): 140-142.
  • Gilkerson JR, Love DN, Whalley JM. 1998. Epidemiology of equine herpesvirus abortion: searching for clues to the future. Aust Vet J, 76(10):675-676.
  • Gupta AK, Rattan B, Malik P, Kaur D, Singh BK, Yadav MP. 2000. Experimental infection of donkeys with EHV-1 of horse origin-a study. J. Equine Sci, 11(2): 29-33.
  • Gür S, Yapici O. 2008. Equine Herpesvirus type 1 and 4 in Individually Reared Horses in Central and Western Turkey. Acta Vet Brno, 77(4): 609-613.
  • Ishiyama T, Nishimori T, Kato M, Yamada H, Sato K, Sentsui H. 1996. Direct detection of equine herpesvirus DNA in tissues of aborted equine fetuses. Zblt Vet Med (B), 43(10): 639-642.
  • Lawrence GL, Gilkerson J, Love DN, Sabine M, Whalley JM. 1993. Rapid single-step differentiation of equid herpesviruses 1 and 4 from clinical material using the polymerase chain reaction and virus specific primers. J Virol Methods, 47: 59-72.
  • Matsumura T., Sugiura T., Imagawa H., Fukunaga Y., Kamada M. 1991. Epizoologitical aspects of type 1 and type 4 equine herpesvirus infections among horse populations. J. Vet. Med. Sci., 54(2):207-211.
  • Matsumura Ta, Kirisawa R, Kondo T, Sugita S, Kamada M. 1994. Molecular epizoologitical studies on respiratory and fetal strains of equid herpesvirus-1 isolated in Japan. Equine Infect Dis, 7: 243-249.
  • Matsumura Tb, Yokota S, Imagawa H, Sugiura T, Wada R, Kanemaru T,Nanbu M, Kirisawa R, Kamada M. 1994. Sero- and molecular-epizoologitical studies on equine herpesvirus type 1 (EHV1) ınfection among Race horses: an occurrence of respiratory disease with nervous disorders. J Equine Sci, 5(2): 59-67.
  • Matsumura T, Kondo T, Sugita S, Damiani AM, O’Callaghan DJ, m Imagawa H. 1998. An equine herpesvirus type 1 recombinant with a deletion in the gE and gI genes is avirulent in young horses. Virology, 242: 68-79.
  • Mengi A, Pfeffer M., Meyer H, Östereider N, Kaaden R, Uyanık F. 1996. İzmit bölgesinden izole edilen equine herpes virusunun moleküler karakteri ve Alman EHV-1 viruslarıyla mukayesesi. Tr J Vet Anim Sci, 20: 169-173.
  • Murphy AF, Gibbs JPE, Horzinek CM, Studdert MJ. 1999. Equine Herpesviruses. Veterinary Virology. Academic Press, San Diego, USA, 3th ed., pp 301-325.
  • Newton SA, Knottenbelt DC, Eldridge PR. 2000. Headshaking in horses: possible aetiopathogenesis suggested by the results of diagnostic tests and several treatment regimens used in 20 cases. Equine Vet J, 32(3): 208-216.
  • O’Callaghan D, Osterreider N. 1999. Equine herpesviruses. In: Granoff A, Webster GR (eds). Encyclopedia of Virology. 2. Ed. Academic Press, London, 508-515.
  • O’Keefe JS, Murray A, Wilks CR, Moriarty KM. 1991. Amplification and differentiation of the DNA of an abort genic (type 1) and a respiratory (type 4) strain of equine herpesvirus by the polymerase chain reaction. Res Vet Sci, 50: 349-351.
  • Packiarajah P, Walker C, Gilkerson J, Whalley MJ, Love DN. 1998. Immune responses and protective efficacy of recombinant baculovirus-expressed glycoproteins of equine herpesvirus 1 (EHV-1) gB, gC and gD alone or in combinations in BALB/c mice. Vet Microbiol, 261-278.
  • Robinson EN. 1997. Current Therapy in Equine Medicine. W.B. Saunders Philadelphia, USA, 4th ed., pp 444-447.
  • Sharma PC, Culinane AA, Onions DE, Nicolson L. 1992. Diagnosis of equid herpesviruses -1 and –4 by polymerase chain reaction. Equine Vet J, 24(1): 20-22.
  • Telford EAR, Watson MS, Perry J, Culinane AA, Davison AJ. 1998. The DNA sequence of equine herpesvirus-4. J Gen Virol, 79: 1197-1203.
  • Tsujimura K, Kondo T, Matsumura T, Sugiura T. 2001. Characterization of a gE-deletion mutant (gE) of equine herpes virus 1 (EHV-1) not containing a foreign gene. J Equine Sci, 12 (3): 111.
  • Turan N, Yildirim F, Altan E, Sennazli G, Gurel A, Diallo I, Yilmaz H. 2012. Molecular and pathological investigations of EHV-1 and EHV-4 infections in horses in Turkey. Res Vet Sci, 93(3): 1504-1507.
  • Van Maanen C, Willink DL, Smeenk LAJ, Brinkhof J, Terpstra C. 2000. An equine herpesvirus 1 (EHV1) abortion storm at a riding school. Vet Quart, 22: 83-87.
  • Van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGecoh DJ, Pringle CR, Wickner RB. 2000. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses, 30-41.
  • Walker C, Love DN; Whalley MJ. 1999. Comparision of the pathogenesis of acute equine herpesvirus 1 (EHV-1) infection in the horse and the mouse model: a review. Vet Microbiol, 68(1-2): 3-13.
  • Yasunaga S, Maeda K, Matsumura T, Kondo T, Kai K. 2000. Application of a type-specific Enzyme-Linkerd- Immunoabsorbent Assay for equine herpesvirus types 1 and 4 (EHV-1 and –4) to horse populations inoculated with inactivated EHV-1 vaccine. J Vet Med Sci, 62(7): 687-691.
  • Yılmaz S. 1971. 1969-1970 de domuz böbreği epitel hücre kültüründe izole ve idantifiye edilen Rhinopneumonitis virusu. Etlik Vet Mikrobiyol Derg, 3 (11-12): 55-62.
  • Yılmaz S, Akçora A, Ertürk A, Girgin H, Karaman Z, Carioğlu B, Ali D, Tatar N. 1995. Türkiye Jokey Kulübü İzmit harası safkan İngiliz kısraklarında tesbit edilen ‘EHV1’ (kısrakların virusu abortus) enfeksiyonu, koruyucu aşılamalar ve alınan sonuçlar. Etlik Vet Mikrobiyol Derg, 8(1): 230-237.
  • Yilmaz H, Gurel A, Aktas M, Yildirim F, Bamac OE, Haktanir D, Tekelioglu BK, Gur E, Tarakci EA, Helps C, Richt JA, Turan N. 2018. Clinical Virological and Pathological Investigations on Horses with Neurologic Disorders in Turkey. JEVS, 67: 1-6.
  • Yildirim Y, Yilmaz V, Kirmizigul AH. 2015. Equine herpes virus type 1 (EHV-1) and 4 (EHV-4) infections in horses and donkeys in northeastern Turkey. Iran J Vet Res, 16(4): 341-344.

Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction

Year 2023, Volume: 6 Issue: 4, 705 - 712, 15.10.2023
https://doi.org/10.19127/bshealthscience.1349444

Abstract

The aim of this study was to use polymerase chain reaction (PCR) to detect and differentiate Equine Herpesvirus Type-1 (EHV-1) and Type-4 (EHV-4) viruses within the racehorse population in Türkiye. The diagnostic sensitivity of PCR was also assessed. For this purpose, 98 nasal swab specimens from naturally infected racehorses aged 2 and above, displaying respiratory symptoms suggestive of EHV infections, and 26 aborted fetuses with various organ samples were collected. DNA extraction and PCR analysis were performed on these samples. The study started with PCR optimization using reference strains of EHV-1 (89c25p) and EHV-4 (TH20p) DNA. Finally optimized was completed and reference strains were used for this study. In conclusion, PCR successfully detected and differentiated 7 EHV-1 positive samples from the tissues of the 26 aborted fetuses, as well as one EHV-1 positive and two EHV-4 positive samples from the nasal swabs of the 98 cases. This study represents one of the pioneering works where PCR was firstly employed to detect and differentiate EHV-1 and EHV-4 strains in Türkiye. The study's findings reveal the presence of both EHV-1 and EHV-4 in Türkiye's racehorse population and is among the early reports to identify the existence of EHV-4 using PCR. These findings underscore the circulation of both viruses within the racehorse population. As a result of this study, it has been concluded that the PCR method is a sensitive, cost-effective, and time-saving diagnostic approach for detecting and distinguishing EHV-1 and EHV-4 infections.

Supporting Institution

This study derived from the PhD thesis VFK231T, received support from the Istanbul University Institute of Health

Project Number

Doktora-VFK231T

Thanks

Istanbul University Institute of Health

References

  • Allen GP, Bryans JT. 1986. Molecular epizootiology, pathogenesis, and prophylaxis of equine herpesvirus-1 infections. Prog Vet Microbiol Immun, 2: 78-144.
  • Ataseven VS, Bilge-Dağalp S, Başaran Z, Keskin S 2010. Seroepidemiological studies of equine herpesviruses 1 (EHV-1) and 4 (EHV-4) infections in working horses from the eastern Turkey. Ankara Üniv Vet Fak Derg, 57: 39-42.
  • Ballagi-Pordany A, Klingeborn B, Flensburg J, Belak S. 1990. Equine herpesvirus type1: Detection of viral DNA sequences in aborted fetuses with the polymerase chain reaction. Vet Microbiol, 22: 373-381.
  • Başkaya H, Keskintepe H, Doğuer M, İyigören B, Yılmaz S, Demir A. 1968. An outbreak of equine virus abortion in Turkey. Isolation and identification of rhino pneumonitis virus in cell cultures. A Ü Vet Fak Derg, 15 (3-4): 309-316.
  • Carvalho R, Passos LMF, Martins AS. 2000. Development of a differential multiplex PCR assay for equine herpesvirus 1 and 4 as a diagnostic tool. J Vet Med, 47:351-359.
  • Carvalho R, Oliveira AM, Souza AM, Passos LMF, Martins AS. 2000. Prevalence of equine herpesvirus type 1 latency detected by polymerase chain reaction. Arch Virol, 145: 1773-1787.
  • Crabb BS, Studdert MJ. 1993. 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, 10: 6332-6338.
  • Csellner H, Walker C, Wellington JE, McLure LE, Love DN, Whalley JM. 2000. EHV-1glycoprotein D (EHV-1 gD) is required for virus entry and cell-cell fusion, and an EHV-1 gD deletion mutant induces a protective immune response in mice. Arch Virol, 145: 2371-2385.
  • Damiani AM, Matsumura T, Yokoyama N, Takeshi M, Takahashi E. 2000. A deletion in the gI and gE genes of equine herpesvirus type 4 reduces viral virulence in the natural host and affects virus transmission during cell to cell spread. Virus Res, 67: 189-202.
  • Damiani AM, Matsumura T, Jang HK, Izumiya Y, Milkami T, Takahashi E. 2000. Identification of the products of the equine herpesvirus type 4 gI and gE genes. Arch Virol, 145: 1489-1496.
  • Donaldson MT, Sweeney CR. 1998. Herpesvirus myeloencephalopathy in horses: 11 cases (1982-1996). J Amer Vet Med Assoc, 213 (5): 671-675.
  • Drummer HE, Reynolds A, Studdert MJ, MacPherson CM, Crabb BS. 1995. Application of an equine herpesvirus 1 (EHV-1) type-specific ELISA to the management of an outbreak of EHV1 abortion. Vet Rec, 136: 579-581.
  • Edington N, Welch HM, Griffiths L. 1994. The prevalence of latent equid herpesviruses in the tissues of 40 abattoir horses. Equine Vet J, 26(2): 140-142.
  • Gilkerson JR, Love DN, Whalley JM. 1998. Epidemiology of equine herpesvirus abortion: searching for clues to the future. Aust Vet J, 76(10):675-676.
  • Gupta AK, Rattan B, Malik P, Kaur D, Singh BK, Yadav MP. 2000. Experimental infection of donkeys with EHV-1 of horse origin-a study. J. Equine Sci, 11(2): 29-33.
  • Gür S, Yapici O. 2008. Equine Herpesvirus type 1 and 4 in Individually Reared Horses in Central and Western Turkey. Acta Vet Brno, 77(4): 609-613.
  • Ishiyama T, Nishimori T, Kato M, Yamada H, Sato K, Sentsui H. 1996. Direct detection of equine herpesvirus DNA in tissues of aborted equine fetuses. Zblt Vet Med (B), 43(10): 639-642.
  • Lawrence GL, Gilkerson J, Love DN, Sabine M, Whalley JM. 1993. Rapid single-step differentiation of equid herpesviruses 1 and 4 from clinical material using the polymerase chain reaction and virus specific primers. J Virol Methods, 47: 59-72.
  • Matsumura T., Sugiura T., Imagawa H., Fukunaga Y., Kamada M. 1991. Epizoologitical aspects of type 1 and type 4 equine herpesvirus infections among horse populations. J. Vet. Med. Sci., 54(2):207-211.
  • Matsumura Ta, Kirisawa R, Kondo T, Sugita S, Kamada M. 1994. Molecular epizoologitical studies on respiratory and fetal strains of equid herpesvirus-1 isolated in Japan. Equine Infect Dis, 7: 243-249.
  • Matsumura Tb, Yokota S, Imagawa H, Sugiura T, Wada R, Kanemaru T,Nanbu M, Kirisawa R, Kamada M. 1994. Sero- and molecular-epizoologitical studies on equine herpesvirus type 1 (EHV1) ınfection among Race horses: an occurrence of respiratory disease with nervous disorders. J Equine Sci, 5(2): 59-67.
  • Matsumura T, Kondo T, Sugita S, Damiani AM, O’Callaghan DJ, m Imagawa H. 1998. An equine herpesvirus type 1 recombinant with a deletion in the gE and gI genes is avirulent in young horses. Virology, 242: 68-79.
  • Mengi A, Pfeffer M., Meyer H, Östereider N, Kaaden R, Uyanık F. 1996. İzmit bölgesinden izole edilen equine herpes virusunun moleküler karakteri ve Alman EHV-1 viruslarıyla mukayesesi. Tr J Vet Anim Sci, 20: 169-173.
  • Murphy AF, Gibbs JPE, Horzinek CM, Studdert MJ. 1999. Equine Herpesviruses. Veterinary Virology. Academic Press, San Diego, USA, 3th ed., pp 301-325.
  • Newton SA, Knottenbelt DC, Eldridge PR. 2000. Headshaking in horses: possible aetiopathogenesis suggested by the results of diagnostic tests and several treatment regimens used in 20 cases. Equine Vet J, 32(3): 208-216.
  • O’Callaghan D, Osterreider N. 1999. Equine herpesviruses. In: Granoff A, Webster GR (eds). Encyclopedia of Virology. 2. Ed. Academic Press, London, 508-515.
  • O’Keefe JS, Murray A, Wilks CR, Moriarty KM. 1991. Amplification and differentiation of the DNA of an abort genic (type 1) and a respiratory (type 4) strain of equine herpesvirus by the polymerase chain reaction. Res Vet Sci, 50: 349-351.
  • Packiarajah P, Walker C, Gilkerson J, Whalley MJ, Love DN. 1998. Immune responses and protective efficacy of recombinant baculovirus-expressed glycoproteins of equine herpesvirus 1 (EHV-1) gB, gC and gD alone or in combinations in BALB/c mice. Vet Microbiol, 261-278.
  • Robinson EN. 1997. Current Therapy in Equine Medicine. W.B. Saunders Philadelphia, USA, 4th ed., pp 444-447.
  • Sharma PC, Culinane AA, Onions DE, Nicolson L. 1992. Diagnosis of equid herpesviruses -1 and –4 by polymerase chain reaction. Equine Vet J, 24(1): 20-22.
  • Telford EAR, Watson MS, Perry J, Culinane AA, Davison AJ. 1998. The DNA sequence of equine herpesvirus-4. J Gen Virol, 79: 1197-1203.
  • Tsujimura K, Kondo T, Matsumura T, Sugiura T. 2001. Characterization of a gE-deletion mutant (gE) of equine herpes virus 1 (EHV-1) not containing a foreign gene. J Equine Sci, 12 (3): 111.
  • Turan N, Yildirim F, Altan E, Sennazli G, Gurel A, Diallo I, Yilmaz H. 2012. Molecular and pathological investigations of EHV-1 and EHV-4 infections in horses in Turkey. Res Vet Sci, 93(3): 1504-1507.
  • Van Maanen C, Willink DL, Smeenk LAJ, Brinkhof J, Terpstra C. 2000. An equine herpesvirus 1 (EHV1) abortion storm at a riding school. Vet Quart, 22: 83-87.
  • Van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGecoh DJ, Pringle CR, Wickner RB. 2000. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses, 30-41.
  • Walker C, Love DN; Whalley MJ. 1999. Comparision of the pathogenesis of acute equine herpesvirus 1 (EHV-1) infection in the horse and the mouse model: a review. Vet Microbiol, 68(1-2): 3-13.
  • Yasunaga S, Maeda K, Matsumura T, Kondo T, Kai K. 2000. Application of a type-specific Enzyme-Linkerd- Immunoabsorbent Assay for equine herpesvirus types 1 and 4 (EHV-1 and –4) to horse populations inoculated with inactivated EHV-1 vaccine. J Vet Med Sci, 62(7): 687-691.
  • Yılmaz S. 1971. 1969-1970 de domuz böbreği epitel hücre kültüründe izole ve idantifiye edilen Rhinopneumonitis virusu. Etlik Vet Mikrobiyol Derg, 3 (11-12): 55-62.
  • Yılmaz S, Akçora A, Ertürk A, Girgin H, Karaman Z, Carioğlu B, Ali D, Tatar N. 1995. Türkiye Jokey Kulübü İzmit harası safkan İngiliz kısraklarında tesbit edilen ‘EHV1’ (kısrakların virusu abortus) enfeksiyonu, koruyucu aşılamalar ve alınan sonuçlar. Etlik Vet Mikrobiyol Derg, 8(1): 230-237.
  • Yilmaz H, Gurel A, Aktas M, Yildirim F, Bamac OE, Haktanir D, Tekelioglu BK, Gur E, Tarakci EA, Helps C, Richt JA, Turan N. 2018. Clinical Virological and Pathological Investigations on Horses with Neurologic Disorders in Turkey. JEVS, 67: 1-6.
  • Yildirim Y, Yilmaz V, Kirmizigul AH. 2015. Equine herpes virus type 1 (EHV-1) and 4 (EHV-4) infections in horses and donkeys in northeastern Turkey. Iran J Vet Res, 16(4): 341-344.
There are 41 citations in total.

Details

Primary Language English
Subjects Epidemiology (Other)
Journal Section Research Article
Authors

Bilge Kaan Tekelioğlu 0000-0001-6727-3175

Ömer Akay This is me 0000-0001-9692-6577

Project Number Doktora-VFK231T
Early Pub Date October 2, 2023
Publication Date October 15, 2023
Submission Date August 27, 2023
Acceptance Date September 29, 2023
Published in Issue Year 2023 Volume: 6 Issue: 4

Cite

APA Tekelioğlu, B. K., & Akay, Ö. (2023). Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction. Black Sea Journal of Health Science, 6(4), 705-712. https://doi.org/10.19127/bshealthscience.1349444
AMA Tekelioğlu BK, Akay Ö. Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction. BSJ Health Sci. October 2023;6(4):705-712. doi:10.19127/bshealthscience.1349444
Chicago Tekelioğlu, Bilge Kaan, and Ömer Akay. “Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses Using Polymerase Chain Reaction”. Black Sea Journal of Health Science 6, no. 4 (October 2023): 705-12. https://doi.org/10.19127/bshealthscience.1349444.
EndNote Tekelioğlu BK, Akay Ö (October 1, 2023) Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction. Black Sea Journal of Health Science 6 4 705–712.
IEEE B. K. Tekelioğlu and Ö. Akay, “Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction”, BSJ Health Sci., vol. 6, no. 4, pp. 705–712, 2023, doi: 10.19127/bshealthscience.1349444.
ISNAD Tekelioğlu, Bilge Kaan - Akay, Ömer. “Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses Using Polymerase Chain Reaction”. Black Sea Journal of Health Science 6/4 (October 2023), 705-712. https://doi.org/10.19127/bshealthscience.1349444.
JAMA Tekelioğlu BK, Akay Ö. Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction. BSJ Health Sci. 2023;6:705–712.
MLA Tekelioğlu, Bilge Kaan and Ömer Akay. “Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses Using Polymerase Chain Reaction”. Black Sea Journal of Health Science, vol. 6, no. 4, 2023, pp. 705-12, doi:10.19127/bshealthscience.1349444.
Vancouver Tekelioğlu BK, Akay Ö. Detecting Equine Herpesvirus-1 and Equine Herpesvirus -4 Infections in Racehorses using Polymerase Chain Reaction. BSJ Health Sci. 2023;6(4):705-12.