1. bookVolume 66 (2016): Issue 4 (December 2016)
Journal Details
License
Format
Journal
eISSN
1820-7448
First Published
25 Mar 2014
Publication timeframe
4 times per year
Languages
English
Open Access

Real-Time PCR Assays for the Specific Detection of Field Balkan Strains of Lumpy Skin Disease Virus

Published Online: 30 Dec 2016
Volume & Issue: Volume 66 (2016) - Issue 4 (December 2016)
Page range: 444 - 454
Received: 10 Nov 2016
Accepted: 06 Dec 2016
Journal Details
License
Format
Journal
eISSN
1820-7448
First Published
25 Mar 2014
Publication timeframe
4 times per year
Languages
English
Abstract

Lumpy skin disease (LSD) is an important disease of cattle which is included in the OIE list of notifiable terrestrial animal diseases because of its great economic importance. The etiological agent is the Lumpy skin disease virus (LSDV).

In the control of LSD attenuated strains of LSDV and SPPV are successfully used as vaccine strains in infected areas. In the case of vaccination policy, due to the possibility of mild or systemic post-vaccination reactions in vaccinated animals, the application of diagnostic procedures that will rapidly and specifically differentiate LSDV field strains from LSD vaccine virus strains are extremely important. Rapidity in diagnostics and disposal of infected animals is one of the key factors in the prevention of spreading the disease.

In the presented study we have described the development and validation of two real-time TaqMan-PCR assays for a rapid, sensitive and specific detection of the virulent field LSDV strain currently circulating in the Balkan Peninsula. Specificity for the field strain and exclusivity for vaccine strains was tested on 171 samples from naturally infected and vaccinated animals.

The results of this study show that both developed real-time PCR assays are more sensitive than the conventional nested PCR in detecting field LSDV strains thus enabling rapid and high-throughput detection of animals infected with field strains of LSDV.

In conclusion, both KV-2 and FLI real-time PCR assays described in this study are simple, rapid, sensitive and suitable for routine use in a diagnostic laboratory and have the potential to replace conventional nested gel-based PCR assays as the standard procedure for the detection of field strains of LSDV in clinical samples.

Keywords

1. Babiuk S, Bowden TR, Parkyn G, Dalman B, Manning L, Neufeld, J. Embury-Hyatt C, Copps J and Boyle DB: Quantification of lumpy skin disease virus following experimental infection in cattle. Transboundary and Emerging Diseases 2008, 55(7):299-307.10.1111/j.1865-1682.2008.01024.xSearch in Google Scholar

2. Babiuk S. Bowden TR. Boyle DB. Wallace DB. Kitching RP: Capripoxviruses: an emerging worldwide threat to sheep, goats and cattle. Transboundary and Emerging Diseases 2008, 55:263-272.10.1111/j.1865-1682.2008.01043.xSearch in Google Scholar

3. Bowden TR, Babiuk SL, Parkyn GR, Copps JS, Boyle DB: Capripoxvirus tissue tropism and shedding: A quantitative study in experimentally infected sheep and goats. Virology 2008, 371(2):380-93.10.1016/j.virol.2007.10.002Search in Google Scholar

4. Carn VM & Kitching RP: The clinical response of cattle experimentally infected with lumpy skin disease (Neethling) virus. Archives of Virology 1995, 140: 503.10.1007/BF01718427Search in Google Scholar

5. International Committee on Taxonomy of Viruses, Virus Taxonomy: 2015 Release, EC 47, London, UK, July 2015.Search in Google Scholar

6. Babiuk S, Bowden TR, Parkyn G, Dalman B, Hoa DM, Long NT, Vu PP, Bieu do X, Copps J, Boyle DB: Yemen and Vietnam capripoxviruses demonstrate a distinct host preference for goats compared with sheep. The Journal of General Virology 2009, 90:105-114.10.1099/vir.0.004507-0Search in Google Scholar

7. Davies FG: Lumpy skin disease, an African Capripoxviruses Disease of Cattle. British Veterinary Journal 1991;147:489-502.10.1016/0007-1935(91)90019-JSearch in Google Scholar

8. Tulman ER, Afonso CL, Lu Z, Zsak L, Kutish GF, Rock DL: Genome of Lumpy Skin Disease Virus. Journal of Virology 2001, 75(15):7122-7130.10.1128/JVI.75.15.7122-7130.2001Search in Google Scholar

9. Tulman ER, Afonso CL, Lu Z, Zsak L, Sur JH, Sandybaev NT, Kerembekova UZ, Zaitsev VL, Kutish GF, Rock DL: The genomes of sheeppox and goatpox viruses. Journal of Virology 2002, 76:6054-6061.10.1128/JVI.76.12.6054-6061.2002Search in Google Scholar

10. Heine HG. Stevens MP. Foord A.J. Boyle DB: A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene. Journal of Immunological Methods 1999. 227:187-196.10.1016/S0022-1759(99)00072-1Search in Google Scholar

11. Haegeman A, Zro K, Vandenbussche F, Demeestere L, Van Campe W, Ennaji MM, De Clercq K: Development and validation of three Capripoxvirus real-time PCRs for parallel testing. J Virol Methods 2013, 193(2):446-51.10.1016/j.jviromet.2013.07.01023850698Search in Google Scholar

12. Hosamani M, Mondal B, Tembhurne PA, Bandyopadhyay SK, Singh RK & Rasool TJ: Differentiation of sheep pox and goat poxviruses by sequence analysis and PCR-RFLP of P32 gene.Virus Genes 2004, 29(1), 73-80.10.1023/B:VIRU.0000032790.16751.13Search in Google Scholar

13. Tuppurainen ES, Venter EH, Coetzer JA: The detection of lumpy skin disease virus in samples of experimentally infected cattle using different diagnostic techniques. The Onderstepoort Journal of Veterinary Research 2005, 72:153-164.10.4102/ojvr.v72i2.213Search in Google Scholar

14. Stram Y, Kuznetzova L, Friedgut O, Gelman B, Yadin H, Rubinstein-Guini M: The use of lumpy skin disease virus genome termini for detection and phylogenetic analysis. J Virol Methods 2008, 151(2):225-9.10.1016/j.jviromet.2008.05.003Search in Google Scholar

15. Lamien CE, Le Goff C. Silber R. Wallace DB, Gulyaz V, Tuppurainen E, Madani H, Caufour P, Adam T, El Harrak M, Luckins AG, Albina E, Diallo A: 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 2011, 149:30-39.10.1016/j.vetmic.2010.09.038Search in Google Scholar

16. Venkatesan G, Balamurugan V, Yogisharadhya R, Kumar A, Bhanuprakash V: Differentiation of sheeppox and goatpox viruses by polymerase Chain reaction-restriction fragment length polymorphism. Virol Sin 2012, 27(6):353-9.10.1007/s12250-012-3277-2Search in Google Scholar

17. Menasherow S, Rubinstein-Giuni M, Kovtunenko A, Eyngor Y, Fridgut O, Rotenberg D, Khinich Y, Stram Y: Development of an assay to differentiate between virulent and vaccine strains of lumpy skin disease virus (LSDV). J Virol Methods 2014, 199:95-101.10.1016/j.jviromet.2013.12.013Search in Google Scholar

18. Menasherow S, Erster O, Rubinstein-Giuni M, Kovtunenko A, Eyngor E, Gelman B, Khinich E, Stram Y: A high-resolution melting (HRM) assay for the differentiation between Israeli fi eld and Neethling vaccine lumpy skin disease viruses. J Virol Methods 2016, 232:12-5.10.1016/j.jviromet.2016.02.008Search in Google Scholar

19. Ireland DC, Binepal YS: Improved detection of capripoxvirus in biopsy samples by PCR. J. Virol. Methods 1998, 74: 1-7.10.1016/S0166-0934(98)00035-4Search in Google Scholar

20. Reed LJ, Muench H: A simple method of estimating fifty per cent endpoints. Am J Hyg 1938, 27:493-497.10.1093/oxfordjournals.aje.a118408Search in Google Scholar

21. Stubbs S, Oura CA, Henstock M, Bowden TR, King DP, Tuppurainen ES: Validation of a high-throughput real-time polymerase chain reaction assay for the detection of capripoxviral DNA. J. Virol. Methods 2012, 179 (2) 419- 422. 10.1016/j.jviromet.2011.11.01522138682Search in Google Scholar

22. Balinsky CA. Delhon G. Smoliga G. Prarat M. French RA. Geary SJ. Rock DL. Rodriguez LL: Rapid preclinical detection of sheeppox virus by a real-time PCR assay. Journal of Clinical Microbiology 2008, 46:438-442.10.1128/JCM.01953-07223812918032617Search in Google Scholar

23. Lamien CE, Lelenta M, Goger W, Silber R, Tuppurainen E, Matijevic M, Luckins AG, Diallo A: Real time PCR method for simultaneous detection, quantitation and differentiation of capripoxviruses. J. Virol. Methods 2011, 171:134-140. 10.1016/j.jviromet.2010.10.01421029751Search in Google Scholar

Recommended articles from Trend MD

Plan your remote conference with Sciendo