Genetic Typing of Bovine Viral Diarrhoea Virus (BVDV) by Restriction Fragment Length Polymorphism (RFLP) and Identification of a New Subtype in Poland

Open access


Restriction fragment length polymorphism (RFLP) analysis was developed for genetic typing of Polish strains of bovine viral diarrhoea virus (BVDV). The method was applied using 60 BVDV isolates, which included BVDV genotype 1, subtypes a, b, d, e, f, and g, and genotype 2a. RT-PCR products of the 5’untranslated region (5’UTR) were digested using three enzymes. Restriction patterns classified the strains into seven groups, each with a specific and different pattern from other subtypes. These findings were confirmed by nucleotide sequencing and phylogenetic analysis. The results suggest that RFLP analysis is a simple, reliable, and fast genotyping method for BVDV strains in comparison with sequencing. This method can distinguish six subtypes of BVDV-1 including a new subtype 1e, identified exclusively by this method, and it allows differentiation of BVDV-1 from BVDV-2 genotype.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • 1. Becher P Orlich M Kosmidou A König M Baroth M Thiel H.J.: Genetic diversity of pestiviruses: identification of novel groups and implications for classification. Virology 1999 262 64-71.

  • 2. Booth R Thomas C.J. El-Attar L.M Gunn G Brownlie J.: A phylogenetic analysis of Bovine Viral Diarrhoea Virus (BVDV) isolates from six different regions of the UK and links to animal movement data. Vet Res 2013 44:43.

  • 3. Buckwold V.E. Beer B.E Donis R.O.: Bovine viral diarrhea virus as a surrogate model of hepatitis C virus for the evaluation of antiviral agents. Antiviral Res 2003 60 1-15.

  • 4. Czopowicz M Kaba J. Schirrmeier H. Bagnicka E Szaluś-Jordanow O. Nowicki M Witkowski L. Frymus T.: Serological evidence for BVDV-1 infection in goats in Poland. Acta Vet Hung 2011 59 399-404.

  • 5. Damen M. Sillekens P Sjerps P Melsert R. Frantzen I Reesink H.W Lelie P.N Cuypers H.T.: Stability of hepatitis C RNA during specimen handling and storage prior to NASBA amplification. J Virol Methods 1998 72 175-184.

  • 6. Gao S. Luo J. Du J. Lang Y. Cong G Shao J. Lin T Zhao F Belák S Liu L Chang H Yin H.: Serological and molecular evidence for natural infection of Bactrian camels with multiple subgenotypes of bovine viral diarrhea virus in Western China. Vet Microbiol 2013 163 172-176.

  • 7. Gołębiowski S Sosnowski A. Żuchowska E.: Mucosal disease and viral diarrhoea in the bison. Med Weter 1972 28 406.

  • 8. Halfon P Khiri H. Gerolami V. Bourliere M. Feryn J.M. Reynier P Gauthier A Cartouzou G: Impact of various handling and storage conditions on quantitative detection of hepatitis C virus RNA. J Hepatol 1996 25 307-311.

  • 9. Harpin S Elahi S.M Cornaglia E. Yolken R.H. Elazhary Y.: The 5’-untranslated region sequence of a potential new genotype of bovine viral diarrhea virus. Arch Virol 1995 140 1285-1290.

  • 10. Hofmann M.A Brechtbühl K. Stäuber N.: Rapid characterization of new pestivirus strains by direct sequencing of PCR-amplified cDNA from the 5’ noncoding region. Arch Virol 1994 139 217-229.

  • 11. Houe H.: Epidemiological features and economical importance of bovine virus diarrhoea virus (BVDV) infections. Vet Microbiol 1999 64 89-107.

  • 12. Houe H.: Economic impact of BVDV infection in dairies. Biologicals 2003 31 137-143.

  • 13. Jackova A Novackova M Pelletier C Audeval C. Gueneau E. Haffar A Petit E Rehby L Vilcek S.: The extended genetic diversity of BVDV-1: typing of BVDV isolates from France. Vet Res Commun 2008 32 7-11.

  • 14. Kuta A. Polak M.P. Larska M. Żmudziński J.F.: Monitoring of bovine viral diarrhoea virus (BVDV) infection in Polish dairy herds using bulk tank milk samples. Bull Vet Inst Pulawy 2013 57 149-156.

  • 15. Kuta A. Polak M.P Larska M. Żmudziński J.F.: Predominance of bovine viral diarrhea virus 1b and 1d subtypes during eight years of survey in Poland. Vet Microbiol 2013 166 639-644.

  • 16. Nagai M. Hayashi M. Itou M. Fakutomi T Akashi H. Kida H. Sakoda Y.: Identification of new genetic subtypes of bovine viral diarrhea virus genotype 1 isolated in Japan. Virus Genes 2008 36 135-139.

  • 17. Neill J.D.: Molecular biology of bovine viral diarrhea virus. Biologicals 2013 41 2-7.

  • 18. Olafson P. McCallum A Fox F.: An apparently new transmissible disease of cattle. Cornell Vet 1946 36 205-213.

  • 19. Parchariyanon S Inui K Pinyochon W Damrongwatanapokin S Takahashi E.: Genetic grouping of classical swine fever virus by restriction fragment length polymorphism of the E2 gene. J Virol Methods 2000 87 145-149.

  • 20. Paton D.J Sands J.J Lowings J.P. Smith J.E Ibata G Edwards S.: A proposed division of the pestivirus genus using monoclonal antibodies supported by cross-neutralisation assays and genetic sequencing. Vet Res 1995 26 92-109.

  • 21. Plowright W.: Other virus diseases in relation to the J.P. 15 programme. In: Joint Campaign Against Rinderpest. First Technical Review Meeting Phase IV. Mogadiscio Kenya 1969 pp. 19-23.

  • 22. Rola M Polak M.P. Rola J. Bicka L Żmudziński J.F. Kuźmak J.: Bovine immunodeficiency virus (BIV) infection in respect to BHV-1 BLV and BVDV coinfections. Med Weter 2005 61 286-289.

  • 23. Rypuła K Kaba J Płoneczko-Janeczko K Wojewoda-Kotwica B Mazurkiewicz M.: Epidemiological aspects of BVDV infections in dairy cattle herds in Poland. Med Weter 2010 66 684-687.

  • 24. Seki Y Seimiya Y.M Motokawa M. Yaegashi G. Nagai M Hayashi M.: Application of restriction fragment length polymorphism analysis to simple and rapid genotyping of bovine viral diarrhea virus strains isolated in Japan. 2008 70 393-395.

  • 25. Tajima M Frey H.R. Yamato O Maede Y. Moennig V Scholz H. Greiser-Wilke I.: Prevalence of genotypes 1 and 2 of bovine viral diarrhea virus in Lower Saxony Germany. Virus Res 2001 76 31-42.

  • 26. Vilcek S. Belák S.: Genetic identification of pestivirus strain Frijters as a border disease virus from pigs. J Virol Methods 1996 60 103-108.

  • 27. Vilcek S Herring AJ. Herring J.A Nettleton P.F Lowings J.P. Paton D.J.: Pestivirus isolated from pigs cattle and sheep can be allocated into at least three genogroups using polymerase chain reaction and restriction endonuclease analysis. Arch Virol 1994 136 309-323.

  • 28. Vilcek S. Paton D.J Durkovic B Strojny L Ibata G Moussa A Loitsch A Rossmanith W Vega S Scicluna M.T Paifi V.: Bovine viral diarrhoea virus genotype 1 can be separated into at least eleven genetic groups. Arch Virol 2001 146 99-115.

  • 29. Vincze T Posfai J Roberts R.J.: NEBcutter: a program to cleave DNA with restriction enzymes. Nucleic Acids Res 2003 31 3688-3691.

Journal information
Impact Factor

IMPACT FACTOR 2018: 0,829
5-year IMPACT FACTOR: 0,938

CiteScore 2018: 0.68

SCImago Journal Rank (SJR) 2018: 0.291
Source Normalized Impact per Paper (SNIP) 2018: 0.501

Cited By
All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 179 82 1
PDF Downloads 91 51 2