Polymerase chain reaction for the differentiation of Marek's disease virus strains

• Autorzy: Kozdrun W , Samorek-Salamonowicz E. , Czekaj H.
• Języki publikacji: EN

Abstrakty

EN

The aim of this study was an attempt to apply PCR for diagnosis of Marek's disease. Eighteen field Marek's disease virus strains and 4 standard strains: virulent HPRS₁₆ strain and attenuated CVI 988 strain (serotype 1 - MDV-1), apathogenic SB 1strain (serotype 2 - MDV-2) and non - pathogenic turkeys HVT FC 126 strain (serotype 3 MDV-3) were used. Chicken anaemia virus (CAV) and infectious laryngotracheitis virus (ILTV) were used as control of PCR. The virus strains were cultured in CEF and total DNA was isolated by A@A Biotechnology kit. Three pairs of primers were used: for gene A of serotype 1, for 132 bp sequence of serotype 1 and for gene A of serotype 3. PCR was carried out under the following conditions: 94°C - 1 min (denaturation), 55°C - 30 s (annealing), 72°C -30 s (elongation). PCR products were analysed by electrophoresis in 2% agarose gel at 100 V/h. The PCR product of 314 bp was obtained from all field strains and HPRS₁₆ standard after the primers for gene A of serotype 1 were used. After the primers for 132 bp sequence of serotype 1 were used, 434 bp fragment for 19 DNA samples derivied from field strains and standard HPRS₁₆ strain and 1033 bp fragment from attenuated CVI 988 strain were obtained. Application of primers for gene A serotype 3 allowed the detection of 436 bp product only from non-pathogenic HVT FC 126 strain (serotype 3). These results indicated that the PCR technique can be used for the differentiation of MDV strains.

Słowa kluczowe

EN

virus strain; Marek's disease; lymphoproliferative disease; diagnosis; DNA isolation; polymerase chain reaction; poultry viral disease; chicken; differentiation

• Wydawca: -
• Czasopismo: Bulletin of the Veterinary Institute in Puławy
• Rocznik: 2001
• Tom: 45
• Numer: 1
• Opis fizyczny: p.5-10,fig.

Twórcy

autor

Kozdrun W

• National Veterinary Research Institute, 24-100 Pulawy, Poland

autor

Samorek-Salamonowicz E.

autor

Czekaj H.

Bibliografia

• 1. Becker Y., Y., Tabor E., Saher Y., Davidson I., Malkinson M., Witter R. L.: PCR detection of amplified 132 bp repeats in Marek's disease virus type 1 (MDV-1) DNA can serve as an indicator for critical genomic rearrangement leading to the attenuation of virus virulence. Virus Gen., 1993, 7, 277 - 287.
• 2. Becker Y., Y., Tabor E., Davidson I., Malkinson M., Weisman Y.: Polymerase chain reaction for differentiation between pathogenic and non-pathogenic serotype 1 Marek's disease viruses (MDV) and vaccine viruses of MDV - serotypes 2 and 3. J. Virol. Meth., 1992, 40, 307 - 322.
• 3. Davidson I., Borovskaya A., Perl S., Malkinson M.: Use of the polymerase chain reaction for the diagnosis of natural infection of chickens and turkeys with Marek's disease virus and reticuloendotheliosis virus. Avian Pathol., 1995, 24, 69 - 94.
• 4. Davidson I., Smith E. J., Perl S., Malkinson M.: Application of the polymerase chain reaction for the diagnosis of natural infection of chickens and turkeys with avian oncogenic viruses. Vet. Med., 1995, 50, 97 - 103.
• 5. Hirai K., Ikuta K., Kato S.: Comparative studies on Marek's disease virus and herpesvirus of turkey DNAs. J. Gen. Virol., 1979, 45, 119 - 131.
• 6. Lee S. F., Kieff E. D., Backenheimer S. L., Roizman B., Spear P. G., Burmester B. R., Nazerian K.: Size and composition of Marek's disease virus deoxyribonucleic acid. J. Virol., 1971, 7, 289 - 294.
• 7. Lee S. Y., Tanaka A., Silver S., Smith M., Nonoyama M.: Minor DNA homology detectable between herpesvirus of turkey and Marek's disease viruses (MDV). Virol., 1979, 93, 277 - 280.
• 8. Mayr A., Bachmann P. A., Bibrack B., Witmann G.: Virusologische Arbeitsmethoden. VEB Gustav Fischer Verlag., 1977, 2, 117 - 121.
• 9. Mullis KB. Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol., 1987, 155, 335-50.
• 10. Rong - Fu W., Beasley J. N., Cao W. W., Slavik M. F., Johnson M.G.: Development of PCR method specific for Marek's disease virus. Mol. Cel. Probes., 1993, 7, 127 - 131.
• 11. Ross L. J. N.: Marek's disease - Molecular biology of the virus. Scientic Basis and Methods of Control. Ed. Matinius Nijhoff Publishing., 1985, 113 - 164.
• 12. Silva R. F.: Differentiation of pathogenic and non-pathogenic serotype 1 Marek's disease viruses (MDVs) by the polymerase chain reaction amplification of the tandem direct repeats within the MDV genome. Avian. Dis., 1992, 36, 521- 528.
• 13. Zerbes M., Tannock G. A., Jenner R. J., Young P. L.: Some characteristics of a recent virulent isolate of Marek's disease virus. Aust. Vet. J., 1994, 71, 21 - 22.
• 14. Zhu G. S., Ojima T., Hironaka T., Ihara T., Mizukoshi N., Kato A., Ueda S., Hirai K.: Differentation of oncogenic and non-oncogenic strain of Marek's disease virus type 1 by using polymerase chain reaction DNA amplification. Avian Dis., 1992, 36, 637 - 645.