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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 3 (march 2019) : 404-407,   Doi: 10.18805/ijar.B-3499

Comparative study on biological material for molecular diagnosis of Gallid herpesvirus 2

V
Venkataramireddy Balena
M
M.R Reddy
R
Rajendra Singh
1Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India
Cite article:- Balena Venkataramireddy, Reddy M.R, Singh Rajendra (2018). Comparative study on biological material for molecular diagnosis of Gallid herpesvirus 2. Indian Journal of Animal Research. 53(3): 404-407. doi: 10.18805/ijar.B-3499.

ABSTRACT

The present study was intended to investigate the sensitivity of different tissues of tumorous and non-tumorous birds to the polymerize chain reaction-baseddiagnosis of Marek’s disease. Tissue samples were collected from spleen, sciatic nerve, feather follicles and tumour parts (if any)of chickens during the routine necropsy examination. The presence of Marek’s disease virus was examined using primers for 132 bp repeats of BamHI-H regionand positive samples were sequenced for 132 bp repeats and Meq gene. In the birds showing tumours, various samples such as the spleen, feather pulp, sciatic nerve and tumour tissue yielded positive results in  92.31%, 88.46%, 100% and 100% cases respectively. Amongst non-tumorous birds, 11.53% spleen, 23.07% feather pulp and 11.53% sciatic nerve were found positive. This is the first comparative study between nerve and other organs samples for molecular diagnosis of Marek’s disease. Data generated shows that the probability of false negative in tumorous birds and false positive in non-tumorous birds was comparatively low while employing nerve tissue than spleen and feather follicles. Thus, it may be concluded from our study that nerves are a more reliable and robust diagnostic organ in comparison to the other organs used for conventional diagnosis.

REFERENCES

  1. Abdul-Careem, M. F., Hunter B. D., Nagy, E., Read, L. R., Sanei, B., Spencer, J. L. and Sharif, S. (2006). Development of a real-time PCR assay using SYBR Green chemistry for monitoring Marek’s disease virus genome load in feather tips. J. Virol. Methods. 133: 34-40.
  2. Abreu, D.L.C., Santos, F.F., José, D.S., Tortelly, R., Nascimento, E.R. and Pereira, V.L.A.(2016). Pathological aspects of a subclinical Marek’s disease case in free-range chickens. Rev. Bras. Cienc. Avic. 18:197-200.
  3. Baigent, S. J., Kgosana, L. B., Gamawa, A. A., Smith, L. P., Read, A. F. and Nair, V. K. (2013). Relationship between levels of very virulent MDV in poultry dust and in feather tips from vaccinated chickens. Avian Dis.57: 440-447.
  4. Baigent, S. J., Petherbridge, L. J., Howes, K., Smith, L. P., Currie, R. J. and Nair, V. K. (2005). Absolute quantitation of Marek’s disease virus genome copy number in chicken feather and lymphocyte samples using real-time PCR. J. Virol. Methods.23: 53–64.
  5. Balachandran, C., Pazhanivel, N., Vairamuthu, S. and Murali Manohar, B.(2009). Marek’s disease and lymphoid leucosis in chicken—    a histopathological survey. Tamilnadu J. Vet. Anim. Sci, 5:167-70.
  6. Bumstead, N., Sillibourne, J., Rennie, M., Ross, N. and Davison, F. (1997). Quantification of Marek’s disease virus in chicken lymphocytes using the polymerase chain reaction with fluorescence detection. J. Virol. Methods.651: 75-81.
  7. Buscaglia, C., Nervi, P. and Risso, M.(2004). Characterization of four very virulent Argentinian strains of Marek’s disease virus and the influence of one of those isolates on synergism between Marek’s disease vaccine viruses. Avian Pathol. 33: 190-195.
  8. Couteaudier, M. and Denesvre, C. (2014).Marek’s disease virus and skin interactions.Vet. res.45: 36.
  9. Davidson, I., Borovskaya, A., Perl, S. and Malkinson, M. (1995).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. : 24:69-94.
  10. Davidson, I. and Borenstein. R. (1999). Multiple infections of chickens and turkeys with avian oncogenic viruses: Prevalence and molecular analysis. ActaVirol.43: 136-142.
  11. Davison, A. J., Eberle, R., Ehlers, B., Hayward, G., McGeoch, D., Minson, A., Pellett, P., Roizman, B., Studdert, M., and Thiry, E. (2009). The order Herpesvirales. Arch. Virol.154: 171-177.
  12. Fodor, I.O.N.I.C.A., Coman, M. and Catana, N. (2001). An outbreak of Marek’s disease in broiler chickens: Epidemiological, clinical and anatomopathological aspects. Lucrari Stiintifice Medicina Veterinaria, 42:129-142.
  13. Islam, A., Cheetham, B. F., Mahony, T. J., Young, P. L. and Walkden-Brown, S. W. (2006). Absolute quantitation of Marek’s disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR. J. Virol.Methods. 132: 127-134.
  14. Luna, L. G. (1968). Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology.
  15. OIE Terrestrial Manual (2010): Chapter 2.3.13.-Marek’s disease.
  16. Sadeghi, M. R., Ghorashi, S. A., Kargar Moakhar, R., Morshedi, D., Salehi Tabar, R., and Ghaemmaghami, S. S. (2006). Polymerase chain reaction for the detection and differentiation of Marek’s disease virus strains MDV-1 and HVT. I J Vet. Res. 7: 17-21.
  17. Sambrook, J. and Russell, D. W. (2001). Detection of DNA in agarose gels. Molecular Cloning: A Laboratory Manual. 1:5-14.
  18. Sambrook, J. and Russell, D.W.(2001). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press.
  19. Schat, A.K. and Nair,V. (2008). Marek’s Disease. In: Diseases of Poultry, Saif, Y.M., A.M. Fadly, J.R. Glisson, L.R. McDougald, L.K. Nolan and D.E. Swayne (Eds.). 12th Edn., Blackwell Publishing, Iowa, pp: 452-512.
  20. Shambhu Dayal Singh, Rajamani Barathidasan, Asok Kumar, Rajib Deb, Amit Kumar Verma and Kuldeep Dhama, (2012). Recent trends in diagnosis and control of Marek’s Disease (MD) in Poultry. Pak J Biol Sci.15: 964-970.
  21. Silva, R. F. (1992). 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., 36: 521-528.
  22. Witter, R. L. (1996). Avian tumor viruses: persistent and evolving pathogens.Acta Vet.Hung. 45: 251-266.
  23. Zelnik, V. (2004). Diagnosis of Marek’s Disease. In: Marek’s Disease: An Evolving Problem, Davison, F. and V. Nair (Eds.). Elsevier Academic Press, London, pp: 157-167. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 3 (march 2019) : 404-407,   Doi: 10.18805/ijar.B-3499

    Comparative study on biological material for molecular diagnosis of Gallid herpesvirus 2

    V
    Venkataramireddy Balena
    M
    M.R Reddy
    R
    Rajendra Singh
    1Division of Pathology, ICAR-Indian Veterinary Research Institute, Izatnagar-243 122, Uttar Pradesh, India
    Cite article:- Balena Venkataramireddy, Reddy M.R, Singh Rajendra (2018). Comparative study on biological material for molecular diagnosis of Gallid herpesvirus 2. Indian Journal of Animal Research. 53(3): 404-407. doi: 10.18805/ijar.B-3499.

    ABSTRACT

    The present study was intended to investigate the sensitivity of different tissues of tumorous and non-tumorous birds to the polymerize chain reaction-baseddiagnosis of Marek’s disease. Tissue samples were collected from spleen, sciatic nerve, feather follicles and tumour parts (if any)of chickens during the routine necropsy examination. The presence of Marek’s disease virus was examined using primers for 132 bp repeats of BamHI-H regionand positive samples were sequenced for 132 bp repeats and Meq gene. In the birds showing tumours, various samples such as the spleen, feather pulp, sciatic nerve and tumour tissue yielded positive results in  92.31%, 88.46%, 100% and 100% cases respectively. Amongst non-tumorous birds, 11.53% spleen, 23.07% feather pulp and 11.53% sciatic nerve were found positive. This is the first comparative study between nerve and other organs samples for molecular diagnosis of Marek’s disease. Data generated shows that the probability of false negative in tumorous birds and false positive in non-tumorous birds was comparatively low while employing nerve tissue than spleen and feather follicles. Thus, it may be concluded from our study that nerves are a more reliable and robust diagnostic organ in comparison to the other organs used for conventional diagnosis.

    REFERENCES

    1. Abdul-Careem, M. F., Hunter B. D., Nagy, E., Read, L. R., Sanei, B., Spencer, J. L. and Sharif, S. (2006). Development of a real-time PCR assay using SYBR Green chemistry for monitoring Marek’s disease virus genome load in feather tips. J. Virol. Methods. 133: 34-40.
    2. Abreu, D.L.C., Santos, F.F., José, D.S., Tortelly, R., Nascimento, E.R. and Pereira, V.L.A.(2016). Pathological aspects of a subclinical Marek’s disease case in free-range chickens. Rev. Bras. Cienc. Avic. 18:197-200.
    3. Baigent, S. J., Kgosana, L. B., Gamawa, A. A., Smith, L. P., Read, A. F. and Nair, V. K. (2013). Relationship between levels of very virulent MDV in poultry dust and in feather tips from vaccinated chickens. Avian Dis.57: 440-447.
    4. Baigent, S. J., Petherbridge, L. J., Howes, K., Smith, L. P., Currie, R. J. and Nair, V. K. (2005). Absolute quantitation of Marek’s disease virus genome copy number in chicken feather and lymphocyte samples using real-time PCR. J. Virol. Methods.23: 53–64.
    5. Balachandran, C., Pazhanivel, N., Vairamuthu, S. and Murali Manohar, B.(2009). Marek’s disease and lymphoid leucosis in chicken—    a histopathological survey. Tamilnadu J. Vet. Anim. Sci, 5:167-70.
    6. Bumstead, N., Sillibourne, J., Rennie, M., Ross, N. and Davison, F. (1997). Quantification of Marek’s disease virus in chicken lymphocytes using the polymerase chain reaction with fluorescence detection. J. Virol. Methods.651: 75-81.
    7. Buscaglia, C., Nervi, P. and Risso, M.(2004). Characterization of four very virulent Argentinian strains of Marek’s disease virus and the influence of one of those isolates on synergism between Marek’s disease vaccine viruses. Avian Pathol. 33: 190-195.
    8. Couteaudier, M. and Denesvre, C. (2014).Marek’s disease virus and skin interactions.Vet. res.45: 36.
    9. Davidson, I., Borovskaya, A., Perl, S. and Malkinson, M. (1995).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. : 24:69-94.
    10. Davidson, I. and Borenstein. R. (1999). Multiple infections of chickens and turkeys with avian oncogenic viruses: Prevalence and molecular analysis. ActaVirol.43: 136-142.
    11. Davison, A. J., Eberle, R., Ehlers, B., Hayward, G., McGeoch, D., Minson, A., Pellett, P., Roizman, B., Studdert, M., and Thiry, E. (2009). The order Herpesvirales. Arch. Virol.154: 171-177.
    12. Fodor, I.O.N.I.C.A., Coman, M. and Catana, N. (2001). An outbreak of Marek’s disease in broiler chickens: Epidemiological, clinical and anatomopathological aspects. Lucrari Stiintifice Medicina Veterinaria, 42:129-142.
    13. Islam, A., Cheetham, B. F., Mahony, T. J., Young, P. L. and Walkden-Brown, S. W. (2006). Absolute quantitation of Marek’s disease virus and Herpesvirus of turkeys in chicken lymphocyte, feather tip and dust samples using real-time PCR. J. Virol.Methods. 132: 127-134.
    14. Luna, L. G. (1968). Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology.
    15. OIE Terrestrial Manual (2010): Chapter 2.3.13.-Marek’s disease.
    16. Sadeghi, M. R., Ghorashi, S. A., Kargar Moakhar, R., Morshedi, D., Salehi Tabar, R., and Ghaemmaghami, S. S. (2006). Polymerase chain reaction for the detection and differentiation of Marek’s disease virus strains MDV-1 and HVT. I J Vet. Res. 7: 17-21.
    17. Sambrook, J. and Russell, D. W. (2001). Detection of DNA in agarose gels. Molecular Cloning: A Laboratory Manual. 1:5-14.
    18. Sambrook, J. and Russell, D.W.(2001). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press.
    19. Schat, A.K. and Nair,V. (2008). Marek’s Disease. In: Diseases of Poultry, Saif, Y.M., A.M. Fadly, J.R. Glisson, L.R. McDougald, L.K. Nolan and D.E. Swayne (Eds.). 12th Edn., Blackwell Publishing, Iowa, pp: 452-512.
    20. Shambhu Dayal Singh, Rajamani Barathidasan, Asok Kumar, Rajib Deb, Amit Kumar Verma and Kuldeep Dhama, (2012). Recent trends in diagnosis and control of Marek’s Disease (MD) in Poultry. Pak J Biol Sci.15: 964-970.
    21. Silva, R. F. (1992). 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., 36: 521-528.
    22. Witter, R. L. (1996). Avian tumor viruses: persistent and evolving pathogens.Acta Vet.Hung. 45: 251-266.
    23. Zelnik, V. (2004). Diagnosis of Marek’s Disease. In: Marek’s Disease: An Evolving Problem, Davison, F. and V. Nair (Eds.). Elsevier Academic Press, London, pp: 157-167. 
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