DEVELOPMENT AND VALIDATION OF A NOVEL DUPLEX PROBE-HYBRIDIZATION QUANTITATIVE PCR FOR LYMPHOMA-ASSOCIATED MIROUNGINE GAMMAHERPESVIRUS 3 IN NORTHERN ELEPHANT SEALS (MIROUNGA ANGUSTIROSTRIS)

J Wildl Dis. 2023 Jan 1;59(1):121-127. doi: 10.7589/JWD-D-22-00030.

Abstract

Recently, a novel gammaherpesvirus, miroungine gammaherpesvirus 3 (MirGHV3), was described in two juvenile elephant seals (Mirounga angustirostris) with diffuse large B-cell lymphoma. We developed and validated a quantitative (q)PCR for rapid detection of MirGHV3 and investigated its potential association with lymphoma. We developed a duplex probe-hybridization qPCR with MirGHV3 DNA polymerase (pol) as the target gene. Each primer-probe combination was cross-validated against the others. Interference was not seen when they were run in the same well as a duplex assay. Twenty-three samples from seven northern elephant seals were tested using the duplex assay. Viral DNA was detected by the assay in 9 of 9 (100%) tissues affected by lymphoma and in 6 of 14 (43%) samples from tissues unaffected by lymphoma. There was a strong correlation between viral copies detected with each of the assays (P=0.0002). Viral load was significantly higher in tissues affected by lymphoma than in those unaffected (P<0.0001). Excluding the virus-negative samples, viral load was still significantly higher in tissues affected by lymphoma than in those unaffected (P=0.0004). This is consistent with a potential role of MirGHV3 in oncogenesis in northern elephant seals, although more studies are needed to determine this definitively. The qPCR developed has utility for further investigations of MirGHV3.

Keywords: Mirounga angustirostris; Elephant seal; gammaherpesvirus; lymphoma; oncogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Viral / isolation & purification
  • Female
  • Gammaherpesvirinae* / genetics
  • Gammaherpesvirinae* / isolation & purification
  • Lymphoma, Large B-Cell, Diffuse* / veterinary
  • Lymphoma, Large B-Cell, Diffuse* / virology
  • Male
  • Polymerase Chain Reaction* / methods
  • Polymerase Chain Reaction* / veterinary
  • Reproducibility of Results
  • Seals, Earless* / virology
  • Tumor Virus Infections* / veterinary
  • Tumor Virus Infections* / virology

Substances

  • DNA, Viral