Skip to main content

Differentiation of sheeppox and goatpox viruses by polymerase Chain reaction-restriction fragment length polymorphism

Virologica Sinica volume 27, pages 352–358 (2012)Cite this article

Abstract

In the present study, the partial gene sequences of P32 protein, an immunogenic envelope protein of Capripoxviruses (CaPV), were analyzed to assess the genetic relationship among sheeppox and goatpox virus isolates, and restriction enzyme specific PCR-RFLP was developed to differentiate CaPV strains. A total of six goatpox virus (GTPV) and nine sheeppox virus (SPPV) isolates of Indian origin were included in the sequence analysis of the attachment gene. The sequence analysis revealed a high degree of sequence identity among all the Indian SPPV and GTPV isolates at both nucleotide and amino acid levels. Phylogenetic analysis showed three distinct clusters of SPPV, GTPV and Lumpy skin disease virus (LSDV) isolates. Further, multiple sequence alignment revealed a unique change at G120A in all GTPV isolates resulting in the formation of Dra I restriction site in lieu of EcoR I, which is present in SPPV isolates studied. This change was unique and exploited to develop restriction enzyme specific PCR-RFLP for detection and differentiation of SPPV and GTPV strains. The optimized PCR-RFLP was validated using a total of fourteen (n=14) cell culture isolates and twenty two (n=22) known clinical samples of CaPV. The Restriction Enzyme specific PCR-RFLP to differentiate both species will allow a rapid differential diagnosis during CaPV outbreaks particularly in mixed flocks of sheep and goats and could be an adjunct/supportive tool for complete gene or virus genome sequencing methods.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

References

  1. Babiuk S, Bowden T R, Boyle D B, et al. 2008. Capripox viruses: An emerging worldwide threat to sheep, goats and cattle. Trans Emerg Dis, 55: 263–272.

    Article  CAS  Google Scholar 

  2. Babiuk S, Bowden T R, Parkyn G, et al. 2009. Yemen and Vietnam capripoxviruses demonstrate a distinct host preference for goats compared with sheep. J Gen Virol, 90(Pt 1):105–114.

    PubMed  Article  CAS  Google Scholar 

  3. Balamurugan V, Danappa Jayappa K, Hosamani M, et al. 2009. Comparative efficacy of conventional and TaqMan polymerase chain reaction assays in the detection of capripoxviruses from clinical samples. J Vet Diagn Invest, 21: 225–231.

    PubMed  Article  Google Scholar 

  4. Balinsky C A, Delhon G, Smoliga G, et al. 2008. Rapid preclinical detection of sheeppox virus by a real-time PCR assay. J Clin Microbiol, 46: 438–442.

    PubMed  Article  CAS  Google Scholar 

  5. Balinsky C A, Delhon G, Afonso C L, et al. 2007. Sheeppox virus kelch-like gene SPPV-019 affects virus virulence. J Virol, 81: 11392–11401.

    PubMed  Article  CAS  Google Scholar 

  6. Bhanuprakash V, Indrani B K, Hosamani M, et al. 2006. The current status of sheeppox disease. Comp Immunol Microbiol Infect Dis, 29: 27–60.

    PubMed  Article  CAS  Google Scholar 

  7. Bhanuprakash V, Venkatesan G, Balamurugan V, et al. 2010. Pox outbreaks in sheep and goats at Makhdoom (Uttar Pradesh), India: evidence of sheeppox virus infection in goats. Transbound Emerg Dis, 57(5): 375–382.

    PubMed  Article  CAS  Google Scholar 

  8. Bhanuprakash V, Hosamani M, Singh R K. 2011. Prospects of control and eradication of capripox from the Indian subcontinent: a perspective. Antiviral Res, 91(3): 225–232.

    PubMed  Article  CAS  Google Scholar 

  9. El-Kenawy A A, El-Tholoth M S, et al. 2010. Sequence analysis of attachment gene of lumpy skin disease and sheep poxviruses. Virol Sin, 25(6): 409–416.

    PubMed  Article  CAS  Google Scholar 

  10. Fulzele S, Singh R K, Hosamani M, et al. 2006. Evaluation of Duplex PCR and PCR-RFLP for Diagnosis of Sheep Pox and Goat Pox. Int J Trop Med, 1(2): 66–70.

    Google Scholar 

  11. Gershon P D, Kitching R P, Hammond J M, et al. 1989. Poxvirus genetic recombination during natural virus transmission. J Gen Virol, 70 (Pt 2): 485–489.

    PubMed  Article  CAS  Google Scholar 

  12. Heine H G, Stevens M P, Foord A J, et al. 1999. A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene. J Immunol Methods, 227: 187–196.

    PubMed  Article  CAS  Google Scholar 

  13. Hosamani M, Mondal B, Tembhurne P A, et al. 2004. Differentiation of sheeppox and goatpox viruses by sequence analysis and PCR-RFLP of P32 gene. Virus Genes, 29: 73–80.

    PubMed  Article  CAS  Google Scholar 

  14. Ireland D C, Binepal Y C. 1998. Improved detection of capripoxvirus in biopsy samples by PCR. J Virol Methods, 74: 1–7.

    PubMed  Article  CAS  Google Scholar 

  15. Lamien C E, Le Goff C, Silber R, et al. 2011. Use of the Capripoxvirus homologue of Vaccinia virus 30 kDa RNA polymerase subunit (RPO30) gene as a novel diagnostic and genotyping target: development of a classical PCR method to differentiate Goat poxvirus from Sheep poxvirus. Vet Microbiol, 149(1–2): 30–39.

    PubMed  Article  CAS  Google Scholar 

  16. Mangana-Vougiouka O, Markoulatos P, Koptopoulos G, et al. 1999. Sheep poxvirus identification by PCR in cell cultures. J Virol Methods, 77 (1):75–79.

    PubMed  Article  CAS  Google Scholar 

  17. Mondal B, Hosamani M, Dutta T K, et al. 2004. An outbreak of sheeppox on a sheep breeding farm in Jammu, India. Rev Sci Tech, 23(3): 943–949.

    PubMed  CAS  Google Scholar 

  18. Parthiban M, Govindarajan R, Manoharan S, et al. 2005. Comparative sequence analysis of diagnostic PCR amplicons from Indian sheeppox virus, Veterinarski Arhiv, 75 (3): 203–209.

    CAS  Google Scholar 

  19. Rao T V S, Bandyopadhyay S K. 2000. A comprehensive review of goatpox and sheeppox and their diagnosis. Anim Health Res Rev, 1(2): 127–136.

    PubMed  Article  CAS  Google Scholar 

  20. Roy P, Purushothaman V, Sreekumar C, et al. 2008. Sheeppox disease outbreaks in Madras Red and Mechery breeds of indigenous sheep in Tamilnadu, India. Res Vet Sci, 85: 617–621.

    PubMed  Article  Google Scholar 

  21. Tamura K, Peterson D, Peterson N, et al. 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Mol Biol Evol, 28(10): 2731–2739.

    PubMed  Article  CAS  Google Scholar 

  22. Van Regenmortel M H V, Fauquet M, Bishop D H L. 2000. Virus taxonomy: Seventh Report of the International Committee on Taxonomy of Viruses. San Diego: Academic Press.

    Google Scholar 

  23. Venkatesan G, Balamurugan V, Singh R K, et al. 2010. Goatpox virus isolated from an outbreak at Akola, Maharashtra (India) phylogenetically related to Chinese strain. Trop Anim Health Prod, 42(6): 1053–1056.

    PubMed  Article  CAS  Google Scholar 

  24. Verma S, Verma L K, Gupta V K, et al. 2011. Emerging Capripoxvirus disease outbreaks in Himachal Pradesh, a northern state of India. Transbound Emerg Dis, 58(1): 79–85.

    PubMed  Article  CAS  Google Scholar 

  25. Yan X M, Chu Y F, Wu G H, et al. 2012. An outbreak of sheeppox associated with goatpox virus in Gansu province of China. Vet Microbiol, 156(3–4): 425–428.

    PubMed  Article  Google Scholar 

  26. Yeruham I, Yadin H, Van Ham M, et al. 2007. Economic and epidemiological aspects of an outbreak of sheeppox in a dairy sheep flock. Vet Rec, 160(7): 236–237.

    PubMed  Article  CAS  Google Scholar 

Download references

Author information

Affiliations

  1. Division of Virology, Indian Veterinary Research Institute, Nainital (Distt.), Mukteswar, 263138, Uttarakhand, India

    Gnanavel Venkatesan, Vinayagamurthy Balamurugan, Revaniah Yogisharadhya, Amit Kumar & Veerakyathappa Bhanuprakash

  2. Project Directorate on Animal Disease Monitoring and Surveillance, H A Farm, Hebbal, Bangalore, 560024, Karnataka, India

    Vinayagamurthy Balamurugan

  3. Indian Veterinary Research Institute, H A Farm, Hebbal, Bangalore, 560024, Karnataka, India

    Veerakyathappa Bhanuprakash

Authors
  1. Gnanavel Venkatesan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Vinayagamurthy Balamurugan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Revaniah Yogisharadhya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Amit Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Veerakyathappa Bhanuprakash
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Veerakyathappa Bhanuprakash.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Venkatesan, G., Balamurugan, V., Yogisharadhya, R. et al. Differentiation of sheeppox and goatpox viruses by polymerase Chain reaction-restriction fragment length polymorphism. Virol. Sin. 27, 352–358 (2012). https://doi.org/10.1007/s12250-012-3277-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12250-012-3277-2

Keywords

  • Sheeppox
  • Goatpox
  • Differential diagnosis
  • PCR-RFLP