Ankara Üniversitesi Veteriner Fakültesi Dergisi
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Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques

Year 2012, Volume: 59 Issue: 1, 35 - 40, 01.03.2012

Volkan YILMAZ Aykut OZKUL

https://doi.org/10.1501/Vetfak_0000002498

Abstract

Bluetongue virus (BTV) is a vector-borne disease of ruminants disseminated in the tropics and sub-tropics. It is also an important problem in the Middle East. The purpose of this study is serotypic identification of local Bluetongue viruses (BTV) isolated between 1998 and 2005. For this purpose, generic (g) and serotype specific (s) RT-PCR systems and the Plaque Reduction Neutralization Assay (PRNA) were used. Generic and serotype specific RT-PCR was performed on the segment 10 and segment 2 levels, respectively. Generic RT-PCR applications revealed specific DNA product (822 bp in length) in all 26 BTV field isolates. On the other hand, 19 BTV isolates were identified as serotype 9 and one isolate was found to be serotype 16, using the sRT-PCR technique. No DNA amplification was observed as a result of sRT-PCR for serotypes 2 and 4. Plaque reduction and neutralization assay (PRNA) was used for serologic identification of BTV isolates. Hyperimmune serums specific to three serotypes (BTV-4, BTV9 and BTV-16) produced in rabbits, were used in PRNA. The test showed that 19 of the isolates were BTV-9, and the remaining two isolates were identified as BTV-4 and BTV-16

Keywords

Bluetongue virus, serotyping identification, PRNA, RT-PCR

References

  • Breard E, Sailleau C, Nomikou K, Hamblin C, Mertens PP, Mellor PS, El Harrak M, Zientara S (2007): Molecular epidemiology of bluetongue virus serotype 4 isolated in Mediterranean Basin between 1979 and 2004. Virus Res, 125, 191–197.
  • Brewer AW, MacLachlan NJ (1992): Ultrastructural characterization of the interaction of bluetongue virus with bovine erythrocytes in vitro. Vet Pathol, 29, 356–359.
  • DeMaula CD, Leutenegger CM, Jutila MA, MacLachlan NJ (2002): Bluetongue virus-induced activation of primary bovine lung microvascular endothelial cells. Vet Immunol Immunopathol, 86, 147–157.
  • Ertürk A (1994): Çeşitli serumlarda (koyun-keçi-sığır) mavi dil antikorlarının agar-jel presipitasyon testi ile araştırılması. Etlik Vet Mikrobiol Derg, 7, 1-19.
  • Frey HR, Liess B (1971): Vermehrungskinetik und verwendbarkeit eines stark zytopathogenen VD-MD virusstammes für diagnostische untersuchungen mit der mikrotiter-methode. Zbl Vet B, 18, 61-71.
  • Gibbs EP, Greiner EC (1994): The epidemiology of bluetongue. Immun Microbiol Infect Dis, 17, 207–220.
  • Gordon SW, Tammariello RF, Linthicum JK, Dohm JD, Digoutte JP, Calvo-Wilson MA (1992): Arbovirus ısolations from mosquitoes collected during 1988 in the Senegal River Basin. Am J Trop Med Hyg, 47, 742-748.
  • Guirakhoo F, Catalan JA, Monath TP (1995): Adaptation of bluetongue virus in mosquito cells results in overexpression of NS3 proteins and release of virus particles. Arch Virol, 140, 967–974.
  • Han Z, Harty RN (2004): The NS3 protein of bluetongue virus exhibits Viroporin-like properties. J Biol Chem, 279, 43092–43097.
  • Hyatt AD, Gould AR, Coupar B, Eaton BT (1991): Localization of the non-structural protein NS3 in bluetongue virus-infected cells. J Gen Virol, 72, 2263– 2267.
  • Karaoğlu T, Özgünlük İ, Demir B, Özkul A, Burgu İ (2007): Seroprevalence of Culicoides-borne disease in cattle in European Turkey. Ankara Üniv Vet Fak Derg, 54, 121-125.
  • Koumbati M, Mangana O, Nomikou K, Mellor PS, Papadopoulos O (1999): Duration of bluetongue viraemia and serological responses in experimentally infected European breeds of sheep and goats. Vet Microbiol, 64, 277–285.
  • Mellor PS, Boorman J (1995): The transmission and geographical spread of African horse sickness and bluetongue viruses. Ann Trop Med Parasitol, 89, 1–15.
  • Mellor PS, Wittmann EJ (2002): Bluetongue virus in the Mediterranean Basin 1998–2001. Vet J, 164, 20–37.
  • Mertens PPC (2004): dsRNA viruses. Virus Res, 101, 3– 13.
  • Mertens PPC, Maan NS, Prasad G, Samuel AR, Shaw AE, Potgieter AC, Anthony SJ, Maan S (2007): Design of primers and use of RT-PCR assays for typing European bluetongue virus isolates: differentiation of field and vaccine strains. J Gen Virol, 88, 2811–2823.
  • Nikolakaki SV, Nomikou K, Koumbati M, Mangana O, Papanastassopoulou M, Mertens PPC, Papadopoulos O (2005): Molecular analysis of the NS3/NS3A gene of Bluetongue virus isolates from the 1979 and 1998–2001 epizootics in Greece and their segregation into two distinct groups. Virus Res, 114, 6–14.
  • OIE (1996): Bluetongue. In: OIE Standards Commission (Ed.), Manual of Standards for Diagnostic Tests and Vaccines. Paris, 109–118.
  • Ozkul A, Yildirim Y, Pinar D, Akçali A, Yilmaz V, Colak D (2006): Serological evidence of West Nile Virus (WNV) in mammalian species in Turkey. Epidemiol Infect, 134, 826-829.
  • Ramakrishnan MA, Pandey AB, Singh KP, Nandi S, Mehrotra ML (2006): Immune responses and protective efficacy of binary ethylenimine (BEI)-Inactivated Bluetongue Virus vaccines in sheep. Vet Rec, 30, 873-880.
  • Roy P (1989): Bluetongue virus proteins. J Gen Virol, 73, 3051-3064.
  • Roy P (2002): Orbivirus. Ed: C. D. Tidona, G. Darai. The Springer Index of Viruses. Springer-Verlag. Berlin, Germany, pp: 957-963.
  • Russel PK, Nisalak A, Sukhavachan P, Vivona S (1967): A plaque reduction test for dengue virus neutralizing antibodies. J Immunol, 99, 285-290.
  • Ulloa A, Langevin SA, Mendez-Sanchez JD, Arredondo-Jimenez JI, Raetz JL, Powers M, Villarreal-Trevino C, Gubler DJ, Komar N (2003): Serologic survey of domestic animals for zoonotic Arbovirus infections in the Lacandón Forest Region of Chiapas, Mexico. Vector-Borne Zoonotic Dis, 3, 3-9.
  • Urman HK, Milli U, Mert N, Berkin S, Kahraman MM, Yüce H, Avvuran H (1979): Türkiye’de buzağılarda konjenital epizootik arthrogryposis ve hydranencephaly olayları. Ankara Üniv Vet Fak Derg, 26, 287-292.
  • Yıldırım Y, Burgu İ (2005): Kuzeydoğu Anadolu Bölgesindeki sığırlarda mavidil (BT), IBR, PI-3, EBL ve BVD enfeksiyonlarının seroprevalansı. Ankara Üniv Vet Fak Derg, 52, 113-117.
  • Yonguç AD, Taylor WP, Csonton L, Worrall E (1982): Bluetongue in western Turkey. Vet Rec, 111, 144-146.
  • Zientara S, Breard E, Sailleau C (2006): Bluetongue: characterization of virus types by reverse transcription- polymerase chain reaction. Dev Biol (Basel), 126, 187– 196

Yerel Mavidil virusu izolatlarının Plak Redüksiyon Nötralizasyon (PRN) ve Reverz TranskriptazPolimeraz Zincir Reaksiyonu (RT-PZR) teknikleri ile serotipik identifikasyonu

Year 2012, Volume: 59 Issue: 1, 35 - 40, 01.03.2012

Volkan YILMAZ Aykut OZKUL

https://doi.org/10.1501/Vetfak_0000002498

Abstract

Mavidil virusu dünyanın subtropik ve tropik bölgelerinde yaygın olan, ruminantların vektörle bulaşan hastalığıdır. Hastalık özellikle Ortadoğu için önem taşımaktadır. Bu çalısmada 1998-2005 yılları arasında izole edilen yerel BT viruslarının serotipik identifikasyonu amaçlanmıştır. Bu amaç için jenerik ve serotip spesifik RT-PZR sistemleri ile PRN testi kullanıldı. Jenerik RT-PZR ve spesifik RT-PZR sırasıyla segment 10 ve segment 2 düzeyinde gerçekleştirildi. Jenerik RT-PCR uygulamalarında 26 adet BTV izolatının tamamında spesifik DNA ürünü (822 bp. boyunda) elde edildi. Diğer taraftan sRT-PZR uygulaması sonunda ise izolatlardan 19 adedi serotip 9 ve 1 adedi serotip 16 olarak identifiye edildi. Serotip 2 ve 4 için uygulanan spesifik RT-PZR sonunda herhangi bir DNA ürünü elde edilemedi. Plak redüksiyon ve nötralizasyon testi BTV izolatlarının serotipik identifikasyonu için kullanıldı. Tavşanlarda üretilen BTV-4, BTV-9 ve BTV-16’ya spesifik hiperimmun serumlar PRN testinde kullanıldı. PRN testi sonunda 19 izolat BTV-9, 1’er izolat BTV-4 ve BTV-16 olarak identifiye edildi

Keywords

Mavidil virusu, serotipik identifikasyon, PRNT, RT-PZR

References

  • Breard E, Sailleau C, Nomikou K, Hamblin C, Mertens PP, Mellor PS, El Harrak M, Zientara S (2007): Molecular epidemiology of bluetongue virus serotype 4 isolated in Mediterranean Basin between 1979 and 2004. Virus Res, 125, 191–197.
  • Brewer AW, MacLachlan NJ (1992): Ultrastructural characterization of the interaction of bluetongue virus with bovine erythrocytes in vitro. Vet Pathol, 29, 356–359.
  • DeMaula CD, Leutenegger CM, Jutila MA, MacLachlan NJ (2002): Bluetongue virus-induced activation of primary bovine lung microvascular endothelial cells. Vet Immunol Immunopathol, 86, 147–157.
  • Ertürk A (1994): Çeşitli serumlarda (koyun-keçi-sığır) mavi dil antikorlarının agar-jel presipitasyon testi ile araştırılması. Etlik Vet Mikrobiol Derg, 7, 1-19.
  • Frey HR, Liess B (1971): Vermehrungskinetik und verwendbarkeit eines stark zytopathogenen VD-MD virusstammes für diagnostische untersuchungen mit der mikrotiter-methode. Zbl Vet B, 18, 61-71.
  • Gibbs EP, Greiner EC (1994): The epidemiology of bluetongue. Immun Microbiol Infect Dis, 17, 207–220.
  • Gordon SW, Tammariello RF, Linthicum JK, Dohm JD, Digoutte JP, Calvo-Wilson MA (1992): Arbovirus ısolations from mosquitoes collected during 1988 in the Senegal River Basin. Am J Trop Med Hyg, 47, 742-748.
  • Guirakhoo F, Catalan JA, Monath TP (1995): Adaptation of bluetongue virus in mosquito cells results in overexpression of NS3 proteins and release of virus particles. Arch Virol, 140, 967–974.
  • Han Z, Harty RN (2004): The NS3 protein of bluetongue virus exhibits Viroporin-like properties. J Biol Chem, 279, 43092–43097.
  • Hyatt AD, Gould AR, Coupar B, Eaton BT (1991): Localization of the non-structural protein NS3 in bluetongue virus-infected cells. J Gen Virol, 72, 2263– 2267.
  • Karaoğlu T, Özgünlük İ, Demir B, Özkul A, Burgu İ (2007): Seroprevalence of Culicoides-borne disease in cattle in European Turkey. Ankara Üniv Vet Fak Derg, 54, 121-125.
  • Koumbati M, Mangana O, Nomikou K, Mellor PS, Papadopoulos O (1999): Duration of bluetongue viraemia and serological responses in experimentally infected European breeds of sheep and goats. Vet Microbiol, 64, 277–285.
  • Mellor PS, Boorman J (1995): The transmission and geographical spread of African horse sickness and bluetongue viruses. Ann Trop Med Parasitol, 89, 1–15.
  • Mellor PS, Wittmann EJ (2002): Bluetongue virus in the Mediterranean Basin 1998–2001. Vet J, 164, 20–37.
  • Mertens PPC (2004): dsRNA viruses. Virus Res, 101, 3– 13.
  • Mertens PPC, Maan NS, Prasad G, Samuel AR, Shaw AE, Potgieter AC, Anthony SJ, Maan S (2007): Design of primers and use of RT-PCR assays for typing European bluetongue virus isolates: differentiation of field and vaccine strains. J Gen Virol, 88, 2811–2823.
  • Nikolakaki SV, Nomikou K, Koumbati M, Mangana O, Papanastassopoulou M, Mertens PPC, Papadopoulos O (2005): Molecular analysis of the NS3/NS3A gene of Bluetongue virus isolates from the 1979 and 1998–2001 epizootics in Greece and their segregation into two distinct groups. Virus Res, 114, 6–14.
  • OIE (1996): Bluetongue. In: OIE Standards Commission (Ed.), Manual of Standards for Diagnostic Tests and Vaccines. Paris, 109–118.
  • Ozkul A, Yildirim Y, Pinar D, Akçali A, Yilmaz V, Colak D (2006): Serological evidence of West Nile Virus (WNV) in mammalian species in Turkey. Epidemiol Infect, 134, 826-829.
  • Ramakrishnan MA, Pandey AB, Singh KP, Nandi S, Mehrotra ML (2006): Immune responses and protective efficacy of binary ethylenimine (BEI)-Inactivated Bluetongue Virus vaccines in sheep. Vet Rec, 30, 873-880.
  • Roy P (1989): Bluetongue virus proteins. J Gen Virol, 73, 3051-3064.
  • Roy P (2002): Orbivirus. Ed: C. D. Tidona, G. Darai. The Springer Index of Viruses. Springer-Verlag. Berlin, Germany, pp: 957-963.
  • Russel PK, Nisalak A, Sukhavachan P, Vivona S (1967): A plaque reduction test for dengue virus neutralizing antibodies. J Immunol, 99, 285-290.
  • Ulloa A, Langevin SA, Mendez-Sanchez JD, Arredondo-Jimenez JI, Raetz JL, Powers M, Villarreal-Trevino C, Gubler DJ, Komar N (2003): Serologic survey of domestic animals for zoonotic Arbovirus infections in the Lacandón Forest Region of Chiapas, Mexico. Vector-Borne Zoonotic Dis, 3, 3-9.
  • Urman HK, Milli U, Mert N, Berkin S, Kahraman MM, Yüce H, Avvuran H (1979): Türkiye’de buzağılarda konjenital epizootik arthrogryposis ve hydranencephaly olayları. Ankara Üniv Vet Fak Derg, 26, 287-292.
  • Yıldırım Y, Burgu İ (2005): Kuzeydoğu Anadolu Bölgesindeki sığırlarda mavidil (BT), IBR, PI-3, EBL ve BVD enfeksiyonlarının seroprevalansı. Ankara Üniv Vet Fak Derg, 52, 113-117.
  • Yonguç AD, Taylor WP, Csonton L, Worrall E (1982): Bluetongue in western Turkey. Vet Rec, 111, 144-146.
  • Zientara S, Breard E, Sailleau C (2006): Bluetongue: characterization of virus types by reverse transcription- polymerase chain reaction. Dev Biol (Basel), 126, 187– 196

Details

Primary Language English
Subjects Veterinary Surgery
Other ID JA84DS75PR
Journal Section Research Article
Authors

Volkan YILMAZ

Aykut OZKUL

Publication Date March 1, 2012
Published in Issue Year 2012Volume: 59 Issue: 1

Cite

Bibtex @letter to editor { auvfd555873, journal = {Ankara Üniversitesi Veteriner Fakültesi Dergisi}, issn = {1300-0861}, eissn = {1308-2817}, address = {}, publisher = {Ankara University}, year = {2012}, volume = {59}, number = {1}, pages = {35 - 40}, doi = {10.1501/Vetfak\_0000002498}, title = {Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques}, key = {cite}, author = {Yılmaz, Volkan and Ozkul, Aykut} }
APA Yılmaz, V. & Ozkul, A. (2012). Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques . Ankara Üniversitesi Veteriner Fakültesi Dergisi , 59 (1) , 35-40 . DOI: 10.1501/Vetfak_0000002498
MLA Yılmaz, V. , Ozkul, A. "Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques" . Ankara Üniversitesi Veteriner Fakültesi Dergisi 59 (2012 ): 35-40 <http://vetjournal.ankara.edu.tr/en/pub/issue/44728/555873>
Chicago Yılmaz, V. , Ozkul, A. "Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques". Ankara Üniversitesi Veteriner Fakültesi Dergisi 59 (2012 ): 35-40
RIS TY - JOUR T1 - Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques AU - VolkanYılmaz, AykutOzkul Y1 - 2012 PY - 2012 N1 - doi: 10.1501/Vetfak_0000002498 DO - 10.1501/Vetfak_0000002498 T2 - Ankara Üniversitesi Veteriner Fakültesi Dergisi JF - Journal JO - JOR SP - 35 EP - 40 VL - 59 IS - 1 SN - 1300-0861-1308-2817 M3 - doi: 10.1501/Vetfak_0000002498 UR - https://doi.org/10.1501/Vetfak_0000002498 Y2 - 2023 ER -
EndNote %0 Ankara Üniversitesi Veteriner Fakültesi Dergisi Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques %A Volkan Yılmaz , Aykut Ozkul %T Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques %D 2012 %J Ankara Üniversitesi Veteriner Fakültesi Dergisi %P 1300-0861-1308-2817 %V 59 %N 1 %R doi: 10.1501/Vetfak_0000002498 %U 10.1501/Vetfak_0000002498
ISNAD Yılmaz, Volkan , Ozkul, Aykut . "Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques". Ankara Üniversitesi Veteriner Fakültesi Dergisi 59 / 1 (March 2012): 35-40 . https://doi.org/10.1501/Vetfak_0000002498
AMA Yılmaz V. , Ozkul A. Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques. Ankara Univ Vet Fak Derg. 2012; 59(1): 35-40.
Vancouver Yılmaz V. , Ozkul A. Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2012; 59(1): 35-40.
IEEE V. Yılmaz and A. Ozkul , "Serotypic identification of local Bluetongue virus isolates using Plaque Reduction Neutralization (PRN) and Reverse Transcriptase Polymerase Chain Reaction (RTPCR) Techniques", Ankara Üniversitesi Veteriner Fakültesi Dergisi, vol. 59, no. 1, pp. 35-40, Mar. 2012, doi:10.1501/Vetfak_0000002498