Molecular detection and characterization of ovine herpesvirus-2 using heminested PCR in Pakistan

J Vet Sci. 2021 Jul;22(4):e51. doi: 10.4142/jvs.2021.22.e51. Epub 2021 Jun 3.

Abstract

Background: Malignant catarrhal fever (MCF) is a highly fatal lymphoproliferative disease of cattle, deer, bison, water buffalo, and pigs caused by the gamma-herpesviruses alcelaphine herpesvirus-1 (AlHV-1) and ovine herpesvirus-2 (OvHV-2).

Objectives: This study aimed to determine the prevalence of OvHV-2 in sheep, goats, cattle, and buffalo in Rawalpindi and Islamabad, Pakistan, by applying molecular and phylogenetic methods.

Methods: Blood samples were aspirated from sheep (n = 54), goat (n = 50), cattle (n = 46) and buffalo (n= 50) at a slaughterhouse and several farms. The samples were subjected to heminested polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the OvHV-2 POL gene and the OvHV-2 ORF75 tegument protein gene.

Results: The highest percentage of MCF positive samples was in sheep (13%), whereas goat, cattle, and buffalo had lower positive percentages, 11%, 9%, and 6.5%, respectively. Four OvHV-2-positive PCR products obtained from sheep samples were sequenced. The sequences obtained were submitted to the NCBI GenBank database (MK852173 for the POL gene; MK840962, MK852171, and MK852172 for the ORF75 tegument protein gene). Phylogenetic analysis revealed a close similarity of study sequences with those of worldwide samples.

Conclusions: This study is the first cross-sectional study on the prevalence and molecular detection of OvHV-2 in apparently healthy cattle and buffalo that could be carrying OvHV-2 acquired from OvHV-2-positive sheep and goats. The results indicate that OvHV-2 is circulating in Pakistan. Further studies are needed to characterize OvHV-2 and elucidate further its prevalence.

Keywords: DNA Viruses, Nested Polymerase Chain Reaction, Ruminants, Pakistan; Malignant catarrhal fever.

MeSH terms

  • Animals
  • Buffaloes
  • Cattle
  • Cross-Sectional Studies
  • Gammaherpesvirinae / genetics
  • Gammaherpesvirinae / isolation & purification*
  • Goats
  • Herpesviridae Infections / epidemiology
  • Herpesviridae Infections / veterinary*
  • Herpesviridae Infections / virology
  • Livestock / virology*
  • Pakistan / epidemiology
  • Phylogeny
  • Polymerase Chain Reaction / methods*
  • Prevalence
  • Sheep
  • Viral Proteins / genetics

Substances

  • Viral Proteins

Supplementary concepts

  • Ovine gammaherpesvirus 2